Hardware

Great hardware is essential for enhancing the potential of great lab work, more so in our hardware submission. In order to systematically enhance and control the production and extraction of our product from our GMO, we have designed a BioReactor along with the subsequent downstream processing to extract our final product. We have used a combination of continuous and batch fed processes.

We run the BioReactor in a continuous mode, retaining the yeast culture in the stationary phase of its cell life cycle where the production of secondary metabolites is at its maximum. We regulate the pH, dissolved oxygen content, and levels of nutrients in a computerized automatic manner. Since the time scale for cell growth is different from the time scale of downstream processing, we grow the cells continuously and the downstream processing happens in a batch- wise manner. This will lead to the scalability of our project resulting in the industrialization of FearOmone production.

Hardware Evolution:

Our initial idea was to follow a popular approach - deploy the yeast on a paper-based device. We had also planned to expand the idea into a novel saw-tooth design to expand the surface area for yeast growth.Our interaction with the farmers made us realize that the conditions in which the yeast was to be deployed were harsh; a paper based device would not survive long, furthermore, once the yeast scent would die out, the rat might damage the paper leading to GMO release, which is a big NO! NO!

We had to make a better design, which could last in storehouse conditions without risking the release of GMOs. After brainstorming, we came up with TOMCAT (our Topologically OptiMized Click Activated Design), a sealed device which would store yeast spores and release them over solid media without being exposed to the environment, on the click of a button. The pheromone produced would pass through a one way filter and create the Fearomone field which would induce fear responses in the rats. This was expanded to a parallel device which could could store the scent that could be sprayed in granaries.

On interacting with the Food Corporation of India, we came to understand that the deployment strategy that was perfect for the farmers' storehouses would not be implementable in the large granaries maintained by the FCI,the strategy that they primarily used was Fumigation of the grains with chemical rodenticides.

All the workers associated with the FCI, CWC, and Punjab Mandi Board were trained for the above, and employing TOM-CAT would be unsuccessful for 3 reasons:

*The uneducated workers wouldn't be able to use the software “RATTEL”.
*The workers were trained to fumigate the grains with sprays,therefore, our solution must demand minimum changes to pre- existing human resources.
*Since TOM-CAT requires low maintenance, the workers would be rendered unemployed, an unfavorable consequence.

Hence, we had to adapt our solution to the organizational framework of the FCI, CWC, and the Punjab Mandi Board.
Therefore, we planned to “THINK BIG” and extract the Felinine from the yeast and store it in liquid form in High Pressure Tanks, which could replace the Chemical Sprays without causing the slightest change in the organizational framework of the Large Store houses.

TOMCAT -Topologically OptiMized Click Activated Tool

In order to deploy our Genetically Modified Organism (GMO), we had to make sure that our product was robust enough to be deployed in the granaries of farmers, have a reasonable shelf life, and be secure enough not to allow any GMO to escape.

This led to the development of TOMCAT and TOMCAT+

Two 3D printed yeast incubating modules, containing highly nutritious media in the form of a hydrogel and our GMO in spore form in a sealed chamber on the inner walls of the device. The device contains another secondary chamber containing the lysis media. A novel feature in our device is that the spores can stay in the sealed chamber in their dormant state for a very long time, and can be spread on top of the media at any time following a simple click mechanism.

Steps to use TOMCAT:

Note: Further, optimization will be carried out to increase the period over which TOMCAT produces FearOmone.

Note: The minimum period after which the second button must be clicked is 2 days, so that the yeast reaches the stationary phase of its life cycle where the production of the secondary metabolites like Felinine would be at their maximum.

Steps to use TOMCAT+:

BIOREACTOR-

“The only way to turn a project into a product is to industrialize it!”

Since we wanted to create an impact on as many stakeholders as possible, we had to extend our Product Design so that it may smoothly be integrated in the organizational framework of the Large Storehouse, where the chief method for avoiding rat infestation was to fumigate the grains with chemical sprays.

The sprays used by the big corporations are toxic nature, but if we were to mass produce a naturally occurring pheromone, it would not only ensure complete eradication of the “RAT MENACE” from the Large Granaries, but will also ensure absolutely no deterioration of the grain quality.

Since, the method of deployment is the same as the one employed by FCI workers, our Product Design can be smoothly integrated with the present strategies without any demand for new resources.

The BIOREACTOR uses a regular supply of nutrients and works in a Batch-cum-Continuous manner to produce Felinine in large quantities.The first chamber maintains the ideal in-vitro condition for maximal production of Felinine, here the culture is incubated for 3 days as the culture reaches stationary phase around the third day.This chamber runs continuously under aerobic conditions.The biomass is passed through a cross-filtration unit to remove cell media from cell mass.In the secondary chamber, the Wet-Milling unit we maintain anaerobic conditions to prevent oxidation of our product after cell lysis, thereby ensuring controlled storage of our product. Our product along with the cell debris is then passed through an Ultra-Filtration unit which remove cell debris.The filtrate is then separated by solvent extraction and the Felinine is isolated using chromatographic techniques. The collected Felinine can then be deployed in High Pressure Sprays and also be stored in hydrogels.High Pressure Sprays can be used to substitute rodenticide sprays in large grain storage corporations and the hydrogels can be used as delayed release device which would allow for the slow release of FearOmone from the hydrogel, making sure that FearOmone stays in the environment for a long time.

Delayed release device

Team:IISER-Mohali/Hardware