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What is BOD Test?

The BOD test acts as an important criterion for knowing the health of an ecosystem. The
BOD Test of Water is done to know how impure or pure the water body is. The BOD test
procedure involves the collection of the water sample in an air-tight container which is then
kept in incubation for five days at 20 ℃. The sample of water contains microorganisms that
have the function of decomposing organic matter. They utilise oxygen present in the water to
do so. BOD measures the amount of oxygen that gets consumed.

Definition of BOD
BOD calculates the amount of dissolved oxygen that is consumed in a water body containing
microorganisms.
What is a BOD Test?

● BOD stands for Biochemical Oxygen Demand. BOD test is mainly done to check the
pollution level of a water body. It is a good indicator of the level of pollution found in a
water body. If the value of BOD is high, it indicates a high level of infestation of
microorganisms in the water.

● BOD aims at calculating the amount of oxygen that is utilised by microorganisms

while decomposing organic matter. A high BOD means the water is unhealthy.
Objectives of BOD Testing
The following are the objectives of BOD:

● BOD is used to measure the level of pollution in a water body. The level of BOD is
directly proportional to the amount of pollution.
● BOD gives an estimate of the overall health of a water body. If the BOD level is High,
the organisms are getting deficient levels of oxygen, including plants and animals.
● Investigating the BOD levels at different points in the water body can help pinpoint
the source of pollution.
● Wastewater treatment aims at improving the water quality of a particular water body.
By comparing the BOD value of influent and effluents, it becomes easier to estimate
the efficiency of wastewater treatment.
● Frequent BOD tests help companies and other industries to remain compliant with
the pollution levels that are allowed for a water body.
● BOD Testing helps in research on marine ecosystems and how it gets affected by
pollution and other factors.
Principle of BOD Test
BOD mainly aims at calculating the dissolved oxygen levels that are consumed by
microorganisms, while decomposing organic stuff. Water is usually collected from a marine
body like a lake or a river. The sample is collected in a way to make sure that no air enters
the sample water. The initial concentration of the dissolved oxygen is calculated. After that,
the sample is kept in an incubator for 5 days at 20℃. In this period, the microorganisms
decompose the organic matter in the water. The final DO (Dissolved Oxygen) is calculated
after 5 days.
Calculation of BOD
The BOD value is calculated by subtracting the Final DO from the initial DO value. The
formula for calculating BOD is
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BOD= (Initial DO- Final DO) x Dilution Factor / Volume of SampleHere,

Here ,
● DO stands for Diffused oxygen
● Initial DO is the one taken before incubation
● The final DO is the one taken after incubation
● The dilution factor is the ratio of the volume of the sample to the diluted sample.
● The value of BOD is expressed in mg/L which denotes the amount of oxygen
consumed.
BOD Test Procedure
The BOD test involves multiple equipment and supplies which are listed below:

● BOD incubator
● Burette and stand
● 300ml BOD bottles: These bottles have glass toppers which ensure air-tight sealing.
● DO Meter
● Wash bottle
● Conical flask
● Measuring cylinder
● Pipette with an elongated tip
● Gloves and seal starch
● The procedure is carried out in the following manner:
Neutralization of Sample
50 ml of water sample is added to 100 ml of water. The pH is adjusted at 7± 0.2 using
sulfuric acid or sodium hydroxide according to the current pH.

Removal of Chlorine Content

To neutralize the chlorine, add the prescribed volume of sodium sulphite solution to the
sample. The chlorine interferes with the proper measurement of the BOD value. Hence it
needs to be neutralized

Preparation of Phosphate Buffer Solution

The Phosphate buffer solution is prepared by dissolving potassium dihydrogen phosphate,
21.75 grams of dipotassium hydrogen phosphate, disodium hydrogen phosphate, and
ammonium chloride in distilled water. Dilute the solution to a volume of 1 litre.

Alkali-Iodide-Azide Reagent
Dissolve sodium hydroxide and sodium iodide (NaI) in distilled water to make a 1000 ml
solution. Then add and dissolve sodium azide in the solution.

Procedure to Test Dissolved Oxygen

The following steps are carried out to test for DO:

● Manganous Sulphate solution is added to the sample using a pipette tip along with
an alkali-iodide-azide reagent.
● The solutions are kept for some time to allow for their reaction with dissolved oxygen.
● Once the precipitates are formed, sulphuric acid is added to dissolve the precipitates.
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● The BOD sample is transferred to an Erlenmeyer flask. Then it is titrated with

Sodium Thiosulfate solution using a starch indicator till the blue hue fades. The
burette reading is noted.
● Perform a similar procedure to determine the burette reading for the blank sample.
BOD Test Measurement Problems
● There can be several reasons why the accuracy of the BOD result might be affected
such as:
● The water sample which is collected from the source needs to reach the laboratory
as soon as possible. If there is a delay in the analysis or a slight contamination of the
water sample, the BOD test will fail.
● Temperature plays an important criteria in determining the result of the BOD test. If
there is a minor change in the incubation temperature, it might hamper the microbial
activity inside the sample.
● If by any chance, there is an addition of organic matter in the sample, the BOD test
results will come out elevated.
● All samples that are collected need to be stored in air-tight bottles to prevent any
contamination of the sample by atmospheric oxygen.
● In some cases, there might be organic compounds that need more incubation time to
degrade. This can result in a lower value of BOD being computed.
● If there are any toxic compounds present in the water sample, the microbial activity of
the microorganisms is disturbed, thus leading to reduced BOD values.
Importance of Biochemical Oxygen Demand
BOD acts as an important parameter in the measurement of water quality. If the BOD turns
out to be high, it means the water is infested with pollutants and sewage. Knowing the BOD
level helps in making insightful decisions regarding new projects that might have an impact
on the environment and the marine ecosystem. BOD also helps in analyzing the
effectiveness of wastewater treatment plants.

Limitations of the BOD Test

● The various limitations of the BOD Test include:
● The BOD is unable to provide information on the non-biodegradable material present
in the water body.
● The amount of time taken is too long in some situations where a rapid test may be
more suitable.
● The BOD results are very sensitive to temperature fluctuations. A slight change might
affect the microbial activity leading to error in results.
● Sometimes in a single sample, different kinds of microbes are present that utilise
oxygen differently. This can generate false results.
● The water samples sometimes contain inhibitory chemicals that may stop the
microbial activity thus falsifying the BOD test.
● The presence of nitrogenous substances like ammonia or nitrate in the sample can
cause alteration to the microbial activity as these too lead to the consumption of
oxygen.
● The type of pollutants in the water cannot be determined using the BOD test.
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Factors Affecting Biochemical Oxygen Demand

● BOD is affected by the following factors:

● Temperature: Higher temperatures fastens the decomposition process, which leads

to an increase in BOD.
● pH Level: Extreme pH values can affect the metabolic activities of microorganisms,
which, in turn, affects BOD.
● Amount of organic matter: The type and concentration of organic and inorganic
substances in water affect BOD levels.

Conclusion
BOD is an important indicator of water pollution. Measuring and controlling BOD levels are
essential for maintaining the health of aquatic ecosystems. By implementing effective
wastewater treatment practices, adopting sustainable agricultural methods, and regulating
industrial effluent discharge, we can reduce the harmful effects of high BOD levels on the
environment.