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. Vitamins in everyone’s home
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. A great part of our project was dedicated to find a cheap and edible medium in which the microbes could be cultivated at home, and to manufacture a simple and practical way to distribute our engineered yeast and bacteria (See Manufacturing page). We found that a microbial culture could be grown in a very simple medium like potato juice, and that the yeast could be lyophilized, mixed with rice flour and water, and distributed in the form of small cubes.

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. 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.

Safety & Regulations

Choice of Strains

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, and which can be toxic at very high dose. The ß-carotene produced by our yeast though is not toxic, since the body only converts what it need into retinol.
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.

European and Indian Regulations

In order to evaluate the feasibility of our product’s implementation, we researched the european and indian regulations concerning the production and distribution of genetically engineered micro-organisms.
We found that the EU directive 90/219/EC of the EC relative to the contained use of GMO would allow production of this product within the european market. This directive is enforced in each of the EU member’s national regulations.

In India, the FSSIE (Food Safety and Standard Authority in India) told us the safety needed to be established in order for our product to be authorized. Both Dr. A. K. Sharma from the FSSAI and Dr. Sunita Grover the Dairy Microbiology Division at the National Dairy Research Institute advised us to chose micro-organisms that were already present in the fermented foods we were targeting, which we did. These organisms are all in Risk Group 1 (Unlikely to cause human disease.), and have the GRAS status (Generally Considered As Safe).
We also found out that the Indian law currently doesn’t allow GM microorganisms because of the use of antibiotic markers that makes them unsafe to eat. But according to Samir K. Brahmachari, former director of the Council of Scientific and Industrial Research in India, our product has a greater chance to be authorized if there is no more live bacteria in the final form of the dish, that is actually eaten. Since the idli is steamed for 10 to 15 minutes before it is eaten, we made an experiment to assess the presence of yeasts and bacteria in the steamed idli, and found out that the steaming process effectively sterilizes the idli and eliminates the microbes present in the batter.

Continuity

As we wanted people to feel that our product really belong to them, we designed it in the form of an open framework that can be modified and improved easily. Based on precious input from the target population and by the Indian community of Paris, this new chassis is made for being manufactured by factories as well as community labs, so people can be sure they will always keep their independance. It allows to reduce the cost of research and development by abstracting the most technical functions and to easily make organisms that can be homegrown and used for nutrient production. This way we hope that people will feel ownership on the product and trust that it only serves their own interests.