Bacillus subtilis project
Bacillus subtilis project
ACKNOWLEDGEMENT
We acknowledge the efforts of each and every person who has lent his/her invaluable
support in this project. In particular we are indebted to our principal for giving us the
prestigious opportunity to participate in the Senior Bal vigyan 2024-25.
In addition to that, we are extremely grateful towards our teachers Ms. Rajshri Kabra,
Ms. Richa Bajpai, Ms. Nandita Mishra and Ms. Priyanka Patodi for their guidance and
appropriate suggestions at every step of the way.
We also extend a heartfelt gratitude to anyone who helped us for the success of this
project.
Lastly, we'd like to thank our parents, our families and all of the others who put up with
our complications and respected our work by helping us whenever they could.
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INTRODUCTION
➢ The Earth is blessed with an abundance of natural resources such as air, wind, soil,
water, forests, flora, and fauna, which have supported human civilization for
thousands of years. However, with the rapid rise of industrialization and usage of
resources indiscriminately by humans, these resources are seriously threatened.
Vital resources like water, mineral deposits, and fossil fuels are now being
overexploited to such an extent that they could be entirely depleted in the near
future.
➢ Water table depletion is a growing concern in many regions due to factors such as
urbanisation, excessive water extraction, and climate change.
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AIM & UTILITY
We aim to create sustainable and durable material that
minimises environmental impact while enhancing structural
integrity. By combining renewable resources, waste products
and other biotic components, we aim to revolutionise the
particular sector and promote a more sustainable and
economically feasible alternative.
We want to diminish the carbon footprint and pollution brought on by the cement
industry, through sustainable and long-lasting materials, these new materials will be able
to decrease the demand for cement-based products with huge footprints in the terms of
production and transportation. We could then produce materials that are not only
sustainable but also strong and durable.
These innovative materials are designed to allow water to pass through their structure,
promoting groundwater recharge and reducing the amount of runoff that enters storm
drains or water bodies. This can help to alleviate water scarcity, mitigate the risk of
flooding, and improve the overall health of local ecosystems.
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NEED FOR A CHANGE
● A country as large as India has a huge requirement for proper infrastructure, the
government has spent a staggering 23 lakh crore rupees on infrastructure in the
past three years but due to poor construction, environmental factors, and improper
maintenance it leaves much to be desired.
● The lack of proper path management leads to loss of life in tragic accidents,
according to WHO India has the highest number of road traffic accidents with over
150,000 deaths per year due to potholes, out of these 60% are due to poor
infrastructure such as potholes, uneven road surfaces and poor maintenance of
pavements.
● Cemented pavements and roads also pose challenges in the percolation of
rainwater into the soil, which overtime has led to significant depletion of the
groundwater levels. According to the central groundwater board 17% of
groundwater blocks in India are under threat out of these 14% are semi critical and
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5% being critical. This is worrying news considering groundwater is the primary
source for irrigation for 60% of farmers; and the main source of drinking water for
85% of the Indian populace.
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● The day when most people will struggle to afford bare necessities like potable
water, is not very far away. According to the World Bank, India houses 18% of the
world’s population, but has only 4% of the world’s water resources. This leaves
163 million Indians lacking access to drinking water.
● Ground water, while being one of the most crucial resources for civilization, is one
of the fastest depleting sources of water. One of the most efficient ways to
replenish groundwater levels is to provide opportunities for water percolation back
into the soil.
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● India being a swiftly developing country, has a high demand for infrastructure
development to meet its economic growth targets and population growth. Most of
this infrastructure is directed in the field of transportation. It is projected that by
2030, transport is expected to attract over 60 percent of infrastructure investment in
India.
● This growth however comes at a cost, the overexploitation of resources like ores,
sand extracted from river beds or through mining, fossil fuels, etc. This rapid
depletion of resources is also paired with equally fast generation of waste. With
agro-waste being a very concerning, yet lesser-known part of the problem. India
generates approximately 500 million tons of agricultural waste annually.
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ENHANCING OUR KNOWLEDGE
There is no end to knowledge. Therefore, to know more about our topic we:
Conducted Surveys
Experimented to
prove
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OUR SURVEYS:
SURVEY 1
The first survey was to identify whether the common people were aware about the
management of water resources and the status of the water table.
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4. Do you feel any water scarcity in your township during the summer months?
a) Yes
b) No
This survey highlighted the decreasing groundwater level in our city and how it
has been affecting the people.
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SURVEY 2:
1. How often have the pavements of this area been changed in the past 10 years?
Ans) Most of the people answered 5-7 years.
2. What is the cost of maintaining these roads and pavements in your society?
Ans) ₹50,000 to 75,00,000 per annum for 200 m2
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3. What according to you is costlier for the maintenance of pavements and other such
constructions?
a) Raw materials
b) Labour
c) Other/Cannot say
4. Are you aware of the environmental consequences of acquiring these raw materials?
a) Yes
b) No
c) Somewhat aware
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Survey analysis
[CATEGORY
NAME]
[CATEGORY
NAME]
[CATEGORY
NAME]
Over 50% of people termed raw materials the most expensive part of maintainence
Over 30% of people said they were aware of the environmental consequences of aquiring these raw
materials
Over 90% of people said they wanted cheaper, more resource efficient and low maintainence
alternative
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INTERVIEWS
The steppingstone to a
revolutionary tomorrow is
to first analyse the
problem. We interviewed
various experts related to
our topic.
➢ Traditionally pavement bricks are mainly made of cement and some amounts of
other materials are also added. In India alone, approximately 298 million tons of
cement is produced each year to meet the ever-increasing demands for
infrastructure development, which includes laying roads, construction of buildings
or for making houses.
➢I
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e cement on such a large scale, massive amounts of raw materials like limestone,
clay and water are required. Obtaining and processing these materials into cement
is an energy intensive process which uses fossil fuels like coal and petroleum to
heat the limestone.
➢ This industry is also water intensive, accounting for 10% of global industrial water
use. Moreover, the cement industry is responsible for up to 5% of worldwide man-
made CO2 emissions.
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PSEUDO BRICK
MATERIAL REQUIRED:
● Sugarcane bagasse,
● cement
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India is a major producer of wheat. With the large production, comes a large
amount of waste generated. A study has proved that 21 million tons of
wheat, equivalent to the entire production of Australia goes waste in India.
PROCEDURE:
This pseudo-brick allows the water to percolate and act as a perfect medium for
water to reach beneath the roads. The brick has more porosity on account of its
composition being made up of mostly agricultural waste. The bricks also offer
moderate strength.
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Easily
degradable
Drawbacks
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OUR SOLUTION
MICRO-MENDING BRICK
(M~2 BRICK)
.The bacteria researched were Bacillus pseudofirmus, Bacillus halodurans, Bacillus
subtilis and Bacillus Pasteurii, to name a few. They possess certain characteristics that
can be used to make self-healing cement with no human intervention required. By
treating our existing pseudo-brick with such bacteria and adding certain other substances,
we formed a brick which is not only resource efficient, economical, and environment
friendly, but also self-healing. We named this feat, the M~2 brick
B. subtilis B. pseudofirmus
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ABOUT THE BACTERIA
SCIENTIFIC PRINCIPLE
➢ The bacteria will be incorporated in our brick by encapsulation in gelatine or
polyurethane. When a crack develops in the brick, it allows water seepage into the
space and this results in the rupturing of the capsule, which releases the endospores
into the environment. These bacteria get favourable conditions to fill up these
cracks with calcium carbonate, effectively healing the crack.
➢ This process can be completed in time as little as a couple of days if provided with
favourable conditions, making MICP an extremely efficient and time saving
process.
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BIOCHEMISTRY BEHIND HEALING
For the bio-cementation process these bacteria perform the organic chemical reaction
urea hydrolysis.
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UREA HYDROLYSIS
The enzyme urease (urea amidohydrolase) is released by these bacteria under suitable
conditions. This enzyme acts as a catalyst for the hydrolysis of urea (CO(NH₂)₂) present
in the environment, and decomposes it into ammonia (NH₃) and carbon dioxide (CO₂).
ammonia further dissociates into ammonium (NH₄⁺) and hydroxide ions (OH⁻) causing
oxidation of carbon dioxide (CO₂) into carbonate ions (CO₃²⁻). In the presence of calcium
ions (Ca²⁺), the carbonate ions react to form calcium carbonate (CaCO₃), which
precipitates as calcite. The newly formed calcium carbonate is deposited between the
cracks, improving the structural integrity of the brick.
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HOW WE MADE THE M~2 BRICK
Organic Ingredients: The M~2 brick utilises a blend of organic ingredients,
including bagasse, soybean, and wheat waste. These materials are selected
for their sustainability, abundance, and potential contribution to the brick's
structural properties.
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Preparation of bacterial culture: A specific strain of bacteria, such as
Bacillus subtilis, is introduced into the nutrient medium and allowed to
multiply.
Encapsulating bacteria in gelatine capsule Adding capsule while preparing the brick
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Setting Period: The brick is allowed to cure for 48 hours, during
which the bacteria establish themselves within the brick matrix.
➢ When these bacteria are applied to a brick made of bagasse and husk, the
biomineralization process improves its strength, and provides self-healing
capabilities.
➢ The M~2 brick is majorly composed of the same materials as the pseudo-brick but
has been treated with ureolytic bacteria and has a source of urea, which in our case
is cow dung. (It is cheap and easily available in all parts of India. We propose to
use this in our M~2 brick as a source of urea.)
By harnessing the power of renewable resources, waste products and natural bacteria, these
materials contribute to a more environmentally responsible and resilient solution, as
research and development in this field progresses, we can expect even greater advancements
in sustainable building materials and practices.
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EXPERIMENTING TO PROVE
PHYSICAL TESTING
EXPERIMENT 1:
OBJECTIVE: To check the porosity of M~2 Bricks
PROCEDURE:
•Take three containers with 1000 ml of water each.
•Put the three bricks (the cement brick, the pseudo brick and the M~2 brick) in each
container
•Keep it for two hours
•Measure the volume of water left in the three containers
Hence the volume of water absorbed in all the three type bricks can be compared
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CONCLUSION: Our test for porosity was successful as we found out that the material
used to make the M~2 brick acts as a balanced medium for water to reach beneath the
pavements on account of its comparatively.
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EXPERIMENT 2:
OBJECTIVE: To test the strength of M~2.
PROCEDURE: To test the strength of the bricks we applied the pressure with weights
and light vehicles.
CONCLUSION: Upon experimentation we found the brick to be strong with the capacity
to withstand a certain amount of pressure.
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BIOCHEMICAL TESTING
EXPERIMENT 1
THEORY: The calcium ions react with the carbonate ions to form calcium carbonate
(CaCO₃), which precipitates out of the solution. This solid formation can be collected
and measured to determine the rate and amount of calcium carbonate emission or
production.
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PROCEDURE:
After adding the calcium chloride, you should observe a white, chalky precipitate
of calcium carbonate forming in the solution. This shows that the reaction is
occurring.
(Calcium ions from CaCl₂ will react with the carbonate ions produced from the
urea breakdown to form calcium carbonate (CaCO₃), which will precipitate out
as a solid.)
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To measure the amount of calcium carbonate formed, you can filter the solution
through filter paper using a funnel. The calcium carbonate will collect on the paper
as a solid.
By adjusting the concentrations of urea, urease, and calcium ions, you can track
how these changes affect the amount of calcium carbonate produced.
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OBSERVATIONS:
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MYTH BUSTED
Q. How do we know that the bacteria we are using isn’t pathogenic ?
Ans. While some bacteria of the Bacillus family are pathogenic, extensive
research has been conducted on the particular bacteria we are using, and they are
not hazardous to humans, animals or plants in any shape or form. As a matter of
fact, one of the bacteria we are using, Bacillus Subtilis is naturally present in
gastrointestinal tracts of animals
No, these bacteria do not degrade the soil quality. Soil acidification is already a
concern in the country, affecting about 48 million hectares (mha) out of 142 mha
of arable land.
These bacteria performs urea hydrolysis and releases ammonia as one of the
byproducts which can help neutralize the soil pH. So it is actually helpful in
improving the soil quality!
Q. How will these bacteria survive when they are not in use?
The Bacillus species of bacteria are known for their highly adaptive nature. They
are naturally found in a wide range of environmental conditions, which means
they can withstand extremely high temperatures ( add temp), alkalinity and lack
of nutrients. Some species' spores have been reported to survive for thousands of
years under the right conditions. This is because they form endospores, which are
highly resistant to unfavourable environmental conditions. These endospores are
tough, nonreproductive structures which consist of the core, which contains the
DNA, ribosomes and enzymes ; the cortex; and the spore coat. Upon receiving
favourable conditions, these endospores quickly turn into vegetative cells and
perform urea hydrolysis.
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Q. India has extremely varied climatic conditions, including extremely high
temperatures, how will these conditions affect these bacteria ?
The Bacillus species is especially known for their ability to adapt to different
conditions easily. The bacteria we are using can survive harsh climatic conditions
and temperatures reaching up to 50 degree celsius. Moreover, they can also form
endospores, which can protect these from harsh conditions.
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COST EFFECTIVENESS
M-2 brick offers a sustainable and cost-effective alternative to traditional bricks, such as
fired clay bricks and cement blocks. The initial production costs will also be significantly
less due to the usage of waste and natural materials such as eggshells and coconut husk,
the long-term benefits in terms of durability, maintenance, and energy efficiency makes
M-2 bricks a more economical choice over the lifetime of a project.
Cost of making 50 M2 Bricks (4”x4”x8”) = Rs175 (per brick – Rs3.5) (add cost of
sourcing culture of bacteria)
Gelatin: Bulk gelatin can cost around 100-200 rupees per kilogram, so a small portion per
brick could cost about 0.5 rupees.
Bacteria: Culturing bacteria in-house costs around 0.2 rupees per brick. Sourcing
specialised strains might raise this cost, so in-house culturing is likely more cost-
effective.
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Cow Dung: Often inexpensive or even free from local farms. Around 0.3 rupees per brick
for cow dung.
Waste Materials: If locally sourced, low-cost waste (like agricultural or industrial waste)
is used, these might only cost around 0.2 rupees per brick.
Eggshells: Eggshells are typically a by-product and often cost-free or minimal if bought
in bulk. Assume a cost of 0.1 rupees per brick.
Adding these up:
• Gelatin: 0.5 rupees
• Bacteria: 0.2 rupees
• Cow Dung: 0.3 rupees
• Waste: 0.2 rupees
• Eggshells: 0.1 rupees
Hence the cost of making a singular M-2 brick is Rs 3.5.
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OUR SUGGESTION
The implementation of M-2 brick in real-world building structures represents a
groundbreaking advancement with far-reaching implications. Beyond their core function
of self-repair, these innovative materials offer a multitude of potential benefits that can
significantly impact the construction industry and society as a whole.
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FUTURE PROSPECTS
We have given a kick start to this technology but its improvement and implementation is
a team effort. With continuous advancements in technology and development, time will
open up new, better avenues for the M~2 brick. Following are some future applications
we thought of -
Constructing roads and highways: India has over 1.5 lakh kms of highways, they occupy
a vast area that stops water from percolating and require tons of cement but with further
research in this field we can potentially build entire roads and highways using this
technology. This feat would ensure stable and increasing ground water levels and reduce
potholes, while remaining more economical and better for the environment.
Smart self healing sensors: To make the brick even more long lasting,
we can add sensors that detect changes in moisture and pH and then
trigger the bacteria to start urea hydrolysis. This could reduce excessive
bacterial activity and prolong the bacteria’s lifespan.
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BIBLIOGRAPHY
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