Khulna University of Engineering & Technology (KUET), Khulna

CE 3242
Sessional on Environmental Engineering II
Exp. Name: Determination of Biological Oxygen Demand (BOD)

Submitted by
Md. Shahariar Kabir
Roll:1701018
Sec: A
Date of Performance: 10 June, 2021
Department of Civil Engineering
Date of Submission: 17 June, 2021
KUET, Bangladesh.
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1. General Discussion

Biochemical oxygen demand (BOD) is defined as the amount of oxygen required by bacteria in
decomposing organic material in a sample under aerobic condition at 20℃ over a period of 5
days. When biodegradable organic matter/waste is released into a water body, microorganisms
(specially bacteria) feed on the wastes, breaking it down to simpler organic and inorganic
substances. When this decomposition takes place in an aerobic environment, it produces non-
objectionable, stable end products (e.g., CO2, SO4, PO4, and NO3) and in this process draws
down the dissolved oxygen (DO) content of water.

Organic matter + O2 CO2 + H2O + New cells + Stable products (bacteria)

Bacteria
The amount of oxygen required by micro-organisms to oxidize organic wastes aerobically is
called biochemical oxygen demand (BOD).

Principle: The method consists of filling with sample, to overflowing, an airtight bottle of the
specified size and incubating it at the specified temperature for 5 d. Dissolved oxygen is
measured initially and after incubation, and the BOD is computed from the difference between
initial and final DO. Because the initial DO is determined shortly after the dilution is made, all
oxygen uptake occurring after this measurement is included in the BOD measurement.

Samples for BOD analysis may degrade significantly during storage between collection and
analysis, resulting in low BOD values. Minimize reduction of BOD by analyzing sample
promptly or by cooling it to near-freezing temperature during storage. However, even at low
temperature, keep holding time to a minimum. Warm chilled samples to 20 ± 3°C before
analysis.

1) Grab samples—If analysis is begun within 2 h of collection, cold storage is unnecessary. If

analysis is not started within 2 h of sample collection, keep sample at or below 4°C from the
time of collection. Begin analysis within 6 h of collection; when this is not possible because the
sampling site is distant from the laboratory, store at or below 4°C and report length and
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temperature of storage with the results. In no case start analysis more than 24 h after grab
sample collection. When samples are to be used for regulatory purposes make every effort to
deliver samples for analysis within 6 h of collection.

2) Composite samples—Keep samples at or below 4°C during compositing. Limit compositing

period to 24 h. Use the same criteria as for storage of grab samples, starting the measurement of
holding time from end of compositing period. State storage time and conditions as part of the
results.

𝐃𝟏 − 𝑫𝟐
𝐁𝐎𝐃𝟓 𝐨𝐟 𝐝𝐢𝐥𝐢𝐭𝐮𝐭𝐞𝐝 𝐬𝐚𝐦𝐩𝐥𝐞 =
𝑷

𝑊ℎ𝑒𝑟𝑒,

D1 = DO of dilluted sample immediately after preparation, mg/L

D2 = DO of dilluted sample after 5 days incubation at 20℃ , mg/L.

P = decimal volumetric fraction of sample used

Again,
𝑆𝑎𝑚𝑝𝑙𝑒 𝑉𝑜𝑙𝑢𝑚𝑒
𝑃=
𝑆𝑎𝑚𝑝𝑙𝑒 𝑣𝑜𝑙𝑢𝑚𝑒 + 𝐷𝑖𝑙𝑢𝑡𝑖𝑜𝑛 𝑤𝑎𝑡𝑒𝑟 𝑣𝑜𝑙𝑢𝑚𝑒

Interference and limitations: Measurements of oxygen consumed in a 5-d test period (5-d BOD
or BOD5, Section 5210B) used here.

Limitations:

 BOD test measures only biodegradable organics.

 Pre-treatment is needed if the sample contains toxic wastes.
 The effects of nitrifying bacteria should be reduced or eliminated by pre-treatment or
by use of inhabitory agents before the test is applied.
 It is essential to have high concentration of active bacteria present in the sample.
Wastewater.
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2. Apparatus

 300 mL BOD bottle

 Lutron DO meter
 BOD Incubator

3. Reagents

 HACH BOD Nutrient Buffer Pillows

4. Procedure

I. 300 mL BOD bottle was prepared by dilution method, then HACH buffer pillow was
used.
II. Initial DO (𝐷1 ) was measured by using DO meter within the BOD bottle.
III. The BOD bottle was closed carefully by using stopper (e.g. bottle cap) and then the
sample was left for 5 days within the BOD incubator.
IV. After 5 days, the DO of the sample (𝐷2 ) was measured by using DO meter.

5. Calculation

D1 = intial DO = 6.8 mg/L

D2 = DO after 5 days = 2.3 mg/L

Sample volume = 5 mL

Dilution water volume = 295 mL

P = decimal volumetric fraction of sample used

𝑆𝑎𝑚𝑝𝑙𝑒 𝑉𝑜𝑙𝑢𝑚𝑒 5 5
𝑃= = = = 0.0167
𝑆𝑎𝑚𝑝𝑙𝑒 𝑣𝑜𝑙𝑢𝑚𝑒 + 𝐷𝑖𝑙𝑢𝑡𝑖𝑜𝑛 𝑤𝑎𝑡𝑒𝑟 𝑣𝑜𝑙𝑢𝑚𝑒 5 + 295 300
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D1 − 𝐷2 6.8 − 2.3
BOD5 of dilituted sample = = = 270 𝑚𝑔/𝐿
𝑃 0.0167

Result: The amount of BOD of total sample is 270 mg/L.

6. Precision and Bias

The experimental result we obtained is consistent with the theory given in which the Biological
Oxygen Demand (BOD) is getting higher if there are a lot of microorganism in water sample. In
this experiment, we got the BOD 270 mg/L. As we know, Industrial waste BOD range is 100-
10,000 mg/L. So, our sample must be Industrial waste water.

From the results, we can see that the lower dilution factor would increase the amount of BOD.
In the presence of free oxygen, aerobic bacteria use the organic matter found in wastewater as
“food.” The BOD test is an estimate of the “food” available in the sample. The more “food”
presence means more dissolved oxygen (DO) required. The BOD test is used to measure waste
water loads to treatment plants, determine plant efficiency (in terms of BOD removal), and
control plant process. It is also used to determine the effects of on discharge on receiving water.

Microorganisms such as bacteria are responsible for decomposing organic waste water. When
organic matter such as dead plants, leaves, grass clipping, sewage or even food waste is present
in water supply, the bacteria will begin the process of breaking down this waste. When this
happens, much of the available dissolved oxygen is consumed by aerobic bacteria, robbing
other aquatic organisms of the oxygen, they need to live. BOD is a chemical procedure for
determining the rate of uptake of dissolved oxygen by the rate of biological organisms in a body
of water use up oxygen. It is not a precise quantitative test, although it is widely used as an
indication of the quality of water.
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7. Reference

1. U.S. ENVIRONMENTAL PROTECTION AGENCY , OFFICE OF RESEARCH AND DEVELOPMENT. 1986. Method-by-
Method Statistics from Water Pollution (WP) Laboratory Performance Evaluation Studies. Quality Assurance
Branch, Environmental Monitoring and Support Lab., Cincinnati, Ohio.

2.https://civilcrews.com/bod/#:~:text=Limitations%20of%20BOD%20test%20BOD%20test%20measures%20o
nly,of%20inhabitory%20agents%20before%20the%20test%20is%20applied.

8. Bibliography

1.YOUNG, J.C., G.N. MCDERMOTT & D. JENKINS . 1981. Alterations in the BOD procedure for the 15th
edition of Standard Methods for the Examination of Water and Wastewater. J. Water Pollut. Control Fed.
53:1253.