denitrifying the level of water pollution

MICROBES DEPENDENT DENITRIFICATION OF CONTAMINATED WATER
INTRODUCTION:-
This is an eco-friendly and economical project. In this we want to show how to reduce the harmful effects of nitrogen which gets mixed with water by natural hazard or else by man-made hazards. With the help of two types of bacteria like thiobascillus and rod shaped bacteria of bacillus species, we want to reduce the adverse effects of the dissolved nitrogen in the water bodies.
HYPOTHESIS:-
Nitrate-rich water containing denitrifying bacteria will exhibit a faster rate of reduction in nitrate levels.
OBJECTIVE:-
To denitrify nitrogen (Nitrate and nitrite) rich water.
SCIENTIFIC PRINCIPLE INVOLVED:-
Incubation of microbes in different concentration of nitrogen (nitrate and nitrite)
Detection of time taken for denitrification.
Detection of contaminant (nitrate and nitrite) by chemical detection method.
WORKING INVESTIGATION:-
Thiobascillus bacteria denitrifies the contaminated water.
UTILITY:-
This project can be used to reduce the nitrogen contamination of water and increase ecological balance of nutrients in aquatic ecosystem.

OVERVIEW:-
Nitrate contamination

The use of fertilizers to improve soil fertility and enhance plant growth has resulted in the contamination of nitrogen compounds such as ammonia, nitrate and nitride in our rivers, lakes and ocean. During soil erosion or heavy rain, flowing water will wash nitrates into the nearest lakes and rivers. This causes the pollution and contamination of the water.
The presence of nitrates in rivers and lakes encourages the growth of algae in the water. The growth of algae will reduce the levels of dissolved oxygen in the water and this poses a problem for the survival of the fish and other living organisms in the water.

Denitrifying bacteria helps to metabolize nitrogen compounds using enzymes to release nitrogen gas. This is an anaerobic process and will only take place in an environment where there is very little oxygen. It is a common occurrence in deep soil and stagnant water. This process is part of the nitrogen cycle, releasing nitrogen gas back into the atmosphere.
NITRATE
The formation of nitrates is an integral part of the nitrogen cycle in our environment. In moderate amounts, nitrate is a harmless constituent of food and water. Due to its high mobility, nitrate also can leach into groundwater. If people or animals drink water high in nitrate, it may cause methemoglobinemia, an illness found especially in infants. Common sources of nitrate include:
• fertilizers and manure,
• animal feedlots,
• municipal wastewater and sludge,
• septic systems, and
• N-fixation from atmosphere by legumes, bacteria and lightning.
BROWN RING TEST
A common nitrate test, known as the brown ring test can be performed by adding ferrous sulpahte to a solution of a nitrate, then slowly adding concentrated sulphuric acid such that the sulphuric acid forms a layer below the aqueous solution. A brown ring will form at the junction of the two layers, indicating the presence of the nitrate ion. Note that the presence of nitrite ions will interfere with this test.
Health Effect of Nitrates
Human
High nitrate levels in water can cause methemoglobinemia or blue baby syndrome, a condition found especially in infants under six months. The stomach acid of an infant is not as strong as in older children and adults. This causes an increase in bacteria that can readily Nitrite is absorbed in the blood, and hemoglobin (the oxygen-carrying component of blood) is converted to methemoglobin . Methemoglobin does not carry oxygen efficiently. This results in a reduced oxygen supply to vital tissues such as the brain. Methemoglobin in infant blood cannot change back to hemoglobin, which normally occurs in adults. Severe methemoglobinemia can result in brain damage and death.onvert nitrate to nitrite (NO2). Do not let infants drink water that exceeds 10 mg/l NO3-N Pregnant women, adults with reduced stomach acidity, and people deficient in the enzyme that changes methemoglobin back to normal hemoglobin are all susceptible to nitrite-induced methemoglobinemia. The most obvious symptom of methemoglobinemia is a bluish color of the skin, particularly around the eyes and mouth. Other symptoms include headache, dizziness, weakness or difficulty in breathing
Animal
Although there is no enforceable drinking water standard for livestock, do not allow animals to drink water with more than 100 mg/l NO3-N. This is especially true of young animals. They are affected by nitrates the same way as human babies. Older animals may tolerate higher levels. Common symptoms include abdominal pain, diarrhea, muscular weakness or poor coordination. Affected animals will have blood that is a chocolate-brown color. If the problem is diagnosed in time, they can fully recover with a treatment of methylene blue.
BASCILLUS
Bacillus (genus Bacillus), any of a group of rod-shaped, gram-positive, aerobic or (under some conditions) anaerobic bacteria widely found in soil and water. The term bacillus has been applied in a general sense to all cylindrical or rod like bacteria.
Thiobascillus
Thiobacillus ,are strictly aerobic bacteria. All species are respiratory organisms.
Thiobacillus ,are obligate autotrophic organisms, meaning they require inorganic molecules as an electron donor and inorganic carbon (such as carbon dioxide) as a source. They obtain nutrients by oxidizing iron and sulfur with O2.

DENITRIFYING
Denitrification also becomes the key pathway for dissimilative nitrate reduction, which is the process in which nitrates are reduced from the soil, the former being highly toxic for living organisms. Denitrification tends to produce large amounts of by-products.
The most common denitrification process is basically outlined below, with the nitrogen oxides being converted back to gaseous nitrogen (as opposed to that of nitrifying bacteria):
2 NO3- + 10 e- + 12 H+ → N2 + 6 H2O

MATERIALS REQUIRED:-
The materials required for the experiment:

- 1000 milliliters of water from an algae-rich pond
- Two glass containers
- 15ml of thiobascillus bacteria
- One syringe
- Ferrous sulphate & Sulphuric acid(concentrated)

PROCEDURE:-
1. For this experiment, the independent variable is the introduction of denitrifying bacteria. The dependent variable is the concentration of nitrate in the water. This is determined by testing the water using the brown ring test. The constants (control variables) are the size of the water sample, the amount of the denitrifying bacteria present in each water sample and the temperature of the environment, which will remain at room temperature.
2. The beakers are labeled as beaker 1 and beaker 2, 300ml of pond water is measured and poured in the beakers, 15ml of Thiobascillus is added to beaker.
3. The beakers are covered using the plastic wrap. The beakers are left aside for 48-86 hours.
4. A sample from both the beakers are taken after the incubation period.
5. A mixture of Sulphuric acid and ferrous sulphate is taken and poured in both the samples.

OBSERVATION:-
From the above activity it was observed that the sample from beaker 2 showed brown ring in the test tube on addition of the mixture of sulphuric acid and ferrous sulphate is taken, while there was no change in the sample from beaker 1.
ADVANTAGES :-
It helps in preventing severe diseases like blue baby syndrome: a disease rarely found among new born babies.
It helps in reducing the presence of algae and other autotrophs which reduce the amount of oxygen.
This project helps us to increase the longitivity of aquatic habitats.

CONCLUSION :-
From the above activity it was concluded that the brown ring formed was due to the presence of nitrate as there was a formation of ring like structure in the lower part of the beaker.
NAME: 1) SAMPANNA PAHI