Lab Manual Environmental Engineering-II Engr.

Arfa Iqbal

EXPERIMENT NO. 07: - DETERMINATION OF BIOCHEMICAL OXYGEN DEMAND

(BOD) IN WASTEWATER SAMPLE.

INTRODUCTION

Biochemical Oxygen Demand (BOD) is usually defined as the amount of oxygen required by
bacteria to oxidize decomposable organic matter under aerobic conditions. 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.
There are three types of bacteria.
1. Aerobic Bacteria

These are the bacteria which require oxygen for their survival/metabolism.
2. Anaerobic Bacteria

These are the bacteria which do not require oxygen for their survival/metabolism.
3. Facultative Bacteria

These are the bacteria which utilize the oxygen if present but can also survive in the absence of
oxygen.
ENVIRONMENTAL 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. For Design of Treatment Facilities
It is an important test for the design of treatment facilities for wastewater.
3. 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.
4. Efficiency of the Treatment Plants
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.

PREPARATION OF SAMPLES
SAMPLING & 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 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.

REAGENTS
1- Manganous sulfate solution (MnSO4)
2- Alkaline potassium iodide-sodium azide solution
3- Sulfuric acid (H2SO4) concentrated
4- Starch indicator solution
5- Sodium thiosulfate (Na2S2O3 5H2O), 0.025 N
APPARATUS
1- Burette stand
2- 300 mL glass stoppered BOD bottles
3- 500 mL wide-mouthed Erlenmeyer flasks
4- Pipettes with elongated tips and minimum volume of 1.0 mL (+/- 0.1 mL)
5- Pipette bulb
6- 250 mL graduated cylinders
7- Distilled water rinse bottle

PROCEDURE
1- 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.
2- Take 9 BOD bottles – note their numbers and arrange them in three groups.
3- Fill each bottle half with the dilution water ensuring that no air gets mixed with it while filling
as in DO test.
4- Add 2ml of domestic wastewater sample in 1st group of bottles. Add 5ml to the 2nd and 10ml
to the 3rd group of bottles.
5- Fill these bottles completely with the dilution media and stopper such that no air bubbles are
entrapped.
6- Now take out one bottle from each group and determine their DO. This will be the initial DO
i.e. D0.
7- Place the rest of the six bottles with sewage sample in the incubator at 20˚C for 5 days.
8- After 5 days find out the 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).
9- 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.
10- Calculate BODs at 20˚C for the sample using following relationship;

D o  D5
BOD(mg / L )   300
ml of sample
OBSERVATIONS & RESULTS:

BOD TABLE FOR 0 DAYS

Volume of Volume of
Sample Dissolved
Fixed Na2S2O3 Mean DO
Bottle # Added Oxygen
Sample Used
(ml) (ml) (ml) (mg/L) (mg/L)
BOD TABLE FOR 5 DAYS
Volume of Volume of
Sample Dissolved
Fixed Na2S2O3 Mean DO
Bottle # Added Oxygen
Sample Used
(ml) (ml) (ml) (mg/L) (mg/L)

DO DEPLETION TABLE
Dissolved Oxygen DO Depleted
Bottle Sample BOD5   300
DO Depleted Vol. of Sample
# Added D0 D5
(ml) (mg/L) (mg/L) (mg/L)

Mean BOD5 = mg/L

COMMENTS:
Question # 1:
Define BOD?

Question # 2:
Why natural water and tap water cannot be used for preparing dilution in the BOD test?

Question # 3:
What is seeding?