EE-II Lab Report # 06 Section B

EXPERIMENT NO. 06:

To Determine Biochemical Oxygen Demand in Waste Water

 What is BOD:
Biochemical Oxygen Demand (BOD) is usually defined as the amount of oxygen required
by bacteria to oxidize decomposable organic matter under aerobic conditions.
 Introduction:
The Biochemical Oxygen Demand (BOD) test takes 5 days to complete and is performed
using a dissolved oxygen test kit. The BOD level is determined by comparing the DO
level of a water sample taken immediately with the DO level of a water sample that has
been incubated in a dark location for 5 days. The difference between the two DO levels
represents the amount of oxygen required for the decomposition of any organic
material in the sample and is a good approximation of the BOD level.
 Organic matter are food of bacteria.
 End products are CO₂ + H₂O.
 BOD Test:
 5 days are required to complete test at temperature 20®C.
 Living organism are not included in Do.
 Living organism are included in BOD.
 Types Of Bacteria:
There are three types of bacteria.
1. Aerobic Bacteria.
2. Anaerobic Bacteria.
3. Facultative Bacteria.
 Aerobic Bacteria:
These are the bacteria which require oxygen for their
survival/metabolism.
 Anaerobic Bacteria:
These are the bacteria which do not require oxygen for their
survival/metabolism.
 Facultative Bacteria:
These are the bacteria which utilize the oxygen if present but can also
Survive in the absence of oxygen.

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EE-II Lab Report # 06 Section B

 Enviromental Significance:
1. Oxygen Requirement of Wastewater for its Stabilization:
It is the principle test applied to find out the strength of wastewater in
terms of oxygen requirements for its stabilization.
2. Stream Pollution Control:
The BOD test is the major criterion used in the stream pollution control
where organic input must be restricted to maintain desired DO levels.
3. For Design of Treatment Facilities:
It is an important test for the design of treatment facilities for
wastewater.
4. Efficiency of the Treatment Plant:
It is used to evaluate the efficiency of various treatment units.
5. Choice of Treatment Method
It is a factor in the choice of treatment method.
 Bioassay Procedure:
 Natural Water Body ( 20 ®C ).
 Specific PH.
 Specific Pressure.
 Toxic Substance are removed from sample provide that condition to
oxidize to more organic matter.
 Bod Bottle should not contact with atmosphere.
 At 5 days DO difference should be 2 𝑚𝑔⁄𝑙 .
 0.5 𝑚𝑔⁄𝑙 of DO should be remained.
According To scientist:
 Unlimited time required to utilize organic matter.
 At 20 Days (90-95%) organic matter utilize.
 At 5 Days (70-80%) organic matter utilize by bacteria.
Nature of BOD Reaction:

𝒅𝒄
∞C
𝒅𝒕
𝒅𝒄
= KC
𝒅𝒕
Rate of reaction is directly proportional to the amount of
biodegradable organic matter.
𝒂 𝒃 𝟑 𝒂 𝟑
CnHaObNc + ( n + 𝟒 - 𝟐 - 𝟒 𝒄 ) O₂ → nCO₂ + ( - 𝟐 𝒄 ) H₂O + CNH₃
𝟐

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EE-II Lab Report # 06 Section B

 Methods:
 Direct Method ( BOD ≤ 7 𝑚𝑔⁄𝑙 )
 Dilution Method ( BOD > 7 𝑚𝑔⁄𝑙 )
 Direct Method:
In this method we do not provide natural condition
BOD = DO₀ – D0₅ (at 20®C)
 Dilution Method:
In this method we provide natural condition.
𝑑𝑐
∞C
𝑑𝑡
 Freedom from toxic material.
 Favorable PH and atmospheric Pressure.
 Presence of available nutrient element.
 Standard Temperature.
 Presence of Micro-organism.
 Preparation of Sample:
 Sampling and Storage:
Samples for BOD test may degrade significantly during storage between
collection and analysis resulting in low BOD values. The samples should be
analyzed immediately, if not they should be cooled to near freezing
temperature during the storage.
 Seeding:
Seeding is defined as “the introduction of microbial/bacterial culture in the
wastewater sample”. It is necessary to have a population of micro-organisms
capable of oxidizing the biodegradable organic matter in the sample.
 Domestic wastewater and surface waters receiving waste
water discharges contain, adequate microbial population
and may not be seeded.
 Some industrial wastewater, particularly having, high
temperature and extreme pH values may not contain
significant microbial population and therefore require
seeding.
 Preparation of Dilution of Water:
Place desired volume of distilled water in a suitable bottle and add phosphate
buffer, magnesium sulfate, calcium chloride, ferric chloride solutions per liter of the
distilled water. Add appropriate seeding material in the sample. Aerate the dilution
water to saturate it with DO.
 Preparation of Dilution of Waste:
 Organic matter are more.
 Incubation of Bottles.

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EE-II Lab Report # 06 Section B

 Detergent used to clean Bottle.

 Initial DO.
 BOD = DO₀ – DO₅
𝐷𝑂₀−𝐷𝑂₅
 Dilution BOD =
𝑃
𝑚𝑙 𝑝𝑖𝑝𝑝𝑒𝑡𝑡𝑒𝑑
 P =
𝑉𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝐵𝑂𝐷 𝐵𝑜𝑡𝑡𝑙𝑒
 Reagents:
 Manganous sulfate solution (MnSO₄)
 Alkaline potassium iodide-sodium azide solution
 Sulfuric acid (H₂SO₄) concentrated
 Starch indicator solution
 Sodium thiosulfate (Na₂S₂O₃ 5H₂O), 0.025 N
 Chemicals Of dilution of Water:
 1 Liter Distilled Water
 1 ml Phosphate Buffer.
 1 ml Sulphate.
 1 ml Calcium Chloride.
 1 ml Ferric Chloride.
 Aeration → To Provide Oxygen.
 Apparatus:
 Burette stand.
 300 mL glass stoppered BOD bottles.
 500 mL wide-mouthed Erlenmeyer flasks.
 Pipettes with elongated tips and minimum volume of 1.0 mL
(+/- 0.1 mL).
 Pipette bulb.
 250 mL graduated cylinders.
 Distilled water rinse bottle.
 Procedure:
i. First of all it is important to know the amount of sample to be used for the test.
For this purpose, the source of the sample is to be recorded.
ii. Take 9 BOD bottles – note their numbers and arrange them in three groups.
iii. Fill each bottle half with the dilution water ensuring that no air gets mixed with it
while filling as in DO test.
iv. Add 2ml of domestic wastewater sample in 1st group of bottles. Add 5ml to the
2nd and 10ml to the 3rd group of bottles.
v. Fill these bottles completely with the dilution media and stopper such that no air
bubbles are entrapped.

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EE-II Lab Report # 06 Section B

vi. Now take out one bottle from each group and determine their DO. This will be
the initial DO i.e. D0₀.
vii. Place the rest of the six bottles with sewage sample in the incubator at 20˚C for 5
days.
viii. After 5 days find out DO in all the bottles (we take the next reading after 5 days
because after 5 days almost 70–80% of the sample is degraded).
ix. That value of oxygen depletion is considered correct which gives an oxygen
depletion of at least 2 mg/L and which have at least 0.5 mg/L DO after 5 days of
incubation.
x. Calculate BODs at 20˚C for the sample using following relationship.

𝑫𝑶₀−𝑫𝑶₅
Dilution BOD =
𝑷

𝒎𝒍 𝒑𝒊𝒑𝒑𝒆𝒕𝒕𝒆𝒅
P =
𝑽𝒐𝒍𝒖𝒎𝒆 𝒐𝒇 𝑩𝑶𝑫 𝑩𝒐𝒕𝒕𝒍𝒆

 Calculations:

 DO at D₀:
 For Sample 2ml:
A= 13.8
B=8
(𝐴−𝐵)∗0.025𝑁∗8
DO = *1000
𝑚𝑙 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 𝑢𝑠𝑒𝑑
(13.8−8)∗0.025𝑁∗8
DO = *1000
300
D0 = 3.86 mg/L

 For sample 5 ml:

A= 4.4
B=1.1

(𝟒.𝟒−𝟏.𝟏)∗𝟎.𝟎𝟐𝟓𝑵∗𝟖
DO = *1000
𝟑𝟎𝟎
D0 = 2.2 mg/L
 For Bottle 10ml:

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EE-II Lab Report # 06 Section B

A= 28.4
B= 24

(𝟐𝟖.𝟒−𝟐𝟒)∗𝟎.𝟎𝟐𝟓𝑵∗𝟖
DO = *1000
𝟑𝟎𝟎
DO = 2.93 mg/L
 DO at D₅:
 For Sample 2ml:
A= 17.4
B=13.8
(𝐴−𝐵)∗0.025𝑁∗8
DO= *1000
𝑚𝑙 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 𝑢𝑠𝑒𝑑
(𝟏𝟕.𝟒−𝟏𝟑.𝟖)∗𝟎.𝟎𝟐𝟓𝑵∗𝟖
DO = *1000
𝟑𝟎𝟎
DO = 2.4 mg/L
 For Sample 5ml:
A= 7.1
B=4.4
(𝟕.𝟏−𝟒.𝟒)∗𝟎.𝟎𝟐𝟓𝑵∗𝟖
DO = *1000
𝟑𝟎𝟎
DO = 1.8 mg/L
 For Sample 10ml:
A= 31.5
B=28.4
(𝟑𝟏.𝟓−𝟐𝟖.𝟒)∗𝟎.𝟎𝟐𝟓𝑵∗𝟖
DO = *1000
𝟑𝟎𝟎

DO = 2.06 mg/L
 BOD Calculation:
For 2ml Sample:
𝐷𝑂₀−𝐷𝑂₅
Dilution BOD =
𝑃
𝑚𝑙 𝑝𝑖𝑝𝑝𝑒𝑡𝑡𝑒𝑑
P =
𝑉𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝐵𝑂𝐷 𝐵𝑜𝑡𝑡𝑙𝑒
3.86−2.4
Dilution BOD =
𝑃

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EE-II Lab Report # 06 Section B

2
P =
300
3.86−2.4
Dilution BOD = ∗ 300
2

Dilution BOD = 219 mg/l

Volume of Volume
Sample No. Sample of sample DO at D₀ DO at D₅
P BOD
Added
(ml) (ml) (mg/L) (mg/L) (mg/L)
2
1 300 2 3.86 2.4 219
300

5
2 300 5 2.2 1.8 24
300

10
3 300 10 2.93 2.06 26.1
300

 Results and Comments:

The BOD of the wastewater sample of different groups comes to be 219
, 24 & 26.1 mg/L .

 If BOD in water body is greater than 80 mg/L it means that it is polluted.

15-CE-62 Hassan Younas Date: 30-11-2018

EE-II Lab Report # 06 Section B

15-CE-62 Hassan Younas Date: 30-11-2018

EE-II Lab Report # 06 Section B

15-CE-62 Hassan Younas Date: 30-11-2018