Team:Paris Bettencourt/Design
Ferment It Yourself
iGEM Paris-Bettencourt 2O15
- Background
- Design
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Should I talk first about what the problem is (malnutrition in India), or will we talk about it somewhere else?
Introduction
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Manufacturing
The aim of the manufacturing project is to find a homemade way to grow and distribute our strains, easily and cheaply.
Growth of our strains
Distribution of our strains
Once the strains are grown, they have to be distributed to the population.
To answer this problem, we designed small cubes, vector of our engineered strains from the community lab to the user house. These cubes were designed to be easy to make: only few common and cheap ingredients are needed, and the recipe is easy to achieve. The idea is also to be flexible to every community lab means, therefore the recipe is not very strict and can be adapted to what the people have available.
The main ingredient needed is rice flour because the cube is supposed to be used for idli, mainly composed of rice. Therefore the use of rice flour doesn’t change the composition of the idli neither its taste.
The cubes have several advantages: it gives the impression of quantity, so effectiveness, since people won’t believe very small amount of material could solve their deficiency problem. But it’s not too big so it doesn’t give the impression of totally changing their idli recipe and their traditions.Recipe
Ingredients
- 2 cups of rice flour
- 1 cup of liquid solution containing the strains (water, potato juice or rice juice, …)
Method
- Mix the 2 cups of flour with the cup of liquid solution
- The dough obtained should not be too sticky, otherwise, add flour
- It should not be too crumbly neither, otherwise, add water
- Divide the dough in small portion
- Shape each portion to obtain a small cube, about half a centimeter in width
- Let the cubes dry at least 2hours
- You should obtain a small solid cube ready to be used
Distribution & (or?) Availability
Cost and time to create the strain (how would you call that?? Also, I need help to assess the cost and/or DALY)
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Access
An important problem we wanted to tackle with our product is the access Indian people have to sources of vitamins. Indeed, we researched what structures and programs already exist in India, and found that it was a major issue.
One of the most important governmental programs for vitamin supplementation is the Integrated Child Development Services (ICDS) program, which has existed in India since 1975. This program monitors the growth and development of children, and provides supplementary nutrition as well as education and primary health care for children under six and pregnant and lactating mothers. The program is implemented through a network of community-level anganwadi centers (AWC), which provides daily supplements both in the center as well as through take-home rations. Those supplements include doses of vitamin A and tablets of iron and folic acid, in order to prevent xerophtalmia - which can lead to blindness - and anaemia.
However, the National Family Health Survey (NFHS-3) held in 2005-2006 in India reports that only 28% of children received those supplements and health care even though 4/5 of children under six lived in a region covered by an anganwadi center. In the 6 months preceding the survey, only 1/4 of the children between 12-35 months were given vitamin A supplements - and in 9 Indian states, this number falls to less than 20%. The access to ICDS has been improved in recent years, however, although access is still far from being universal (2). According to a national survey held in 2013-14, 46% of children aged 6-59 months had received doses vitamin A.
As for folic acid, the 2013-14 survey reports only 14% of children aged 6-59 months having received iron and folic acid supplements.
Though the program holds great promise and has implemented more than a million anganwadi centers, it is clear that people’s access to them is still very limited. People have to walk to the nearest center everyday if they want to receive the supplements, which is not convenient especially in rural areas.This observation held a major role in our design of a product that can be grown at home, in every village or household. A culture of microorganisms doesn’t require an expensive infrastructure; in fact, most Indian families have already been growing microbial cultures that are several generations old by using a part of their daily yogurt as a starter for the fermentation of the next day’s yogurt. As we have shown (Célia’s part), a microbial culture can be grown on or in a very simple media like potato juice.
We believe that a culture of yeast and bacteria that can be grown at home and supplements fermented foods with vitamins could reach far more people than the anganwadicenters: those cultures would only need to be distributed once, which would not be very costly as the packages can be made to be very small and light and one culture would act as a starter culture for many more. In this way, the culture would be maintained in the home, without the need for people to travel long distances to have access to nutritional supplements.
Our product is also more accessible in that it doesn't have the same geographic and climate constraints as cultures of rice or other crops. Contrary to Golden Rice, the rice developed by the Swiss Federal Institute of Technology and the University of Freiburg that biosynthesizes beta-carotene and can only be grown in environments with very high water availability, a microbial culture can be grown anywhere. Even though this summer we mostly focused on microbial cultures of fermented dishes composed of rice, microbes are used to ferment any kind of cereals as well as other foods. So our product can be adapted to a very wide range of fermented dishes, and could also be used by people who don’t eat rice.
Acceptance
People's opinion
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Why idli?
Idli (or idly) is a small steamed cake made with fermented rice and black lentils (called urad dal or dal). It is a very popular breakfast food across India, mostly in the south, and is often eaten with chutney.
We wanted our product to truly fit the traditions and tastes of the people we were targeting, and idli appeared to be the ideal dish. It is a very popular staple of the Indian diet, along with the crepe-like dosa, which is made of a similar but less coarse batter. Its two ingredients (rice and dal) are very cheap and widely available resources, and idli is commonly eaten by people who lack other types of food and who suffer from vitamin deficiencies. There is even a program that distributes free rice to the poorest populations in the country, the Public Distribution System (PDS); and in the city of Chennai and its suburbs, canteens called Amma Unavagam sell idli and other foods for very low prices (Rs1 for 1 idli).
Besides its popularity and easy accessibility, the most interesting property of idli is that it is fermented. To cook idli, people soak rice and dal separately, then grind and mix them together, then let the batter ferment overnight. In the hot Indian climate, the batter ferments quickly and its volume can triple overnight.
Many studies have been made on the microbiome of idli batter. Though the strains found in idli can be highly variable from one study to another, probably because the microbiome varies from different regions, we found that Lactococcus lactis and Lactobacillus plantarum are among the most common fermentative bacteria found in idli batter, while in the yeast population, Saccharomyces cerevisiae was always present. ADD SOURCES Since we wanted our product to disrupt as little as possible the idli's taste, we chose these organisms for production of the different vitamins and to improve iron availability, instead of adding new micro-organisms that weren’t already present in the idli batter that may have influenced the microbiome in an unpredictable manner. Many people who reviewed our project said that taste was of high importance, and would prefer that we modified microbes already present in the idli rather than add new ones.
+ should we talk about the advantages of fermented foods?Idli recipe
Taste & color
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An orthogonal GMO (is that even english??)
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Safety & Regulations
Choice of Strains
Since our product is meant to be used freely in households without containment, and to be eaten by people, safety is a major element we had to address. In this optic we chose to implement our pathways in Generally Considered As Safe (GRAS) organisms: Saccharomyces cerevisiae, Lactobacillus plantarum, Lactococcus lactis and Propionibacterium freudenreichii are all GRAS, commonly found in food.
We used Escherichia Coli to assemble our plasmids because this bacteria is easy to work with, but our goal is to have the vitamins produced by GRAS only.
A further step to our project would be to implement the pathways in wild type strains directly taken from fermented foods, instead of lab strains. or would it change anything if we did that, in regards to safety and regulations?? I don’t know
Toxicity
During the summer we worked on the synthesis of 3 vitamins: vitamin A, vitamin B2, and vitamin B12.
No toxic or adverse effects have been associated with B12, even in very large intakes, so there is no tolerable upper intake level (UL) over which the B12 becomes toxic. And this is true for all the forms of B12 present in food and supplements.
The same is not true of vitamin A, though: most vitamin pills contain retinol, which is the form of vitamin A that is used by the human body. JB je te laisse écrire c'est quoi qui est toxique et pourquoi la ß-carotene l'est pas !! :)
The vitamin B2 that our micro-organism is producing is riboflavin, which can be toxic at very high dose. However it is very unlikely that we will produce more that the daily requirement (about 1.2mg/day), contrarily to the vitamin pills that we can buy on the market and that often contain a lot more, from 10 to 100 mg. Barth do you want to add/change anything? Please do! :)
European Regulations
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Indian Regulations
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Final Product or Continuity
Someone else (Antoine ? <3) needs to fill that...
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Design
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Note
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