1.

INTRODUCTION
Microorganisms such as bacteria are responsible for decomposing organic waste.

When organic matter such as dead plants, leaves, grass clippings, manure, sewage, or even
food waste is present in a 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.
Biochemical oxygen demand (BOD) or sometimes referred to as Biological oxygen
demand a procedure to determine the amount of dissolved oxygen needed by aerobic
biological organisms in a body of water to break down organic material present in a given
water sample at certain temperature over a specific time period. It is commonly used as the
indicator to show the cleanliness of the waste water.
Regarding waste waters, BOD is often used as a measure of the strength of the waste.
The greater the BOD, the more concentrated the waste. BOD is somewhat unique in that it
measures an impact on the environment (mass of dissolved oxygen consumed per volume of
water sample).
The BOD value is most commonly expressed in milligrams of oxygen consumed per
litre of sample during 5 days of incubation at 20 C and is often used as a robust surrogate of
the degree of organic pollution of water. BOD is amount of oxygen required to decay a
certain amount of organic matter. If too much organic matter is added, the available oxygen
supplies will be used up. 1 mg/L of BOD will, after uptake by bacteria, decrease the DO level
by 1 mg/L.

2.0

OBJECTIVE
The main objective of BOD experiment is to measure the amount of dissolved oxygen

in the waste water for a specified period of time and temperature.

3.0

THEORY
BOD is defined as the amount of oxygen required by living organisms in the

stabilization of the organic matter of water. If the oxidation of an organic compound is carried
1

out by microorganisms using the organic matter as a food source, the oxygen is consumed.
The greater the amount of organic matter present, the greater the amount of oxygen utilized.
The BOD test is indirect measurement of organic matter. Since the test is performed over a
five day period, it is often referred as five day BOD (BOD5).
It is generally assumed that the rate at which oxygen is consumed directly
proportional to the concentration of degradable organic matter remaining at any time.
Different result would be obtained at different temperatures because biochemical reaction
rates are temperature-dependent. The kinetics of the BOD reaction are, for particle purposes,
formulated in accordance with first-order reaction kinetics and may be expressed as
dL / dt = - kLt
where, Lt is the amount of the first stage BOD remaining in the water at time t and k is the
reaction rate constant. This equation can be integrated as
ln Lt / L = - kt
or
The amount of BOD remaining at time t equals
Lt = L e-kt
and y the amount of BOD that has been exerted at any time t, equals
BODt = L( 1 e-kt )
For five day BOD (BOD5), the sample is kept in incubator which remains around
20oc. Most of the biological process spread up as the temperature increase and slow down as
the temperature drop. The temperature of 20oc is standard temperature in lab and it is suitable
for the bacteria active in breaking down the waste.

4.0

MATERIAL AND EQUIPMENT

1. Beaker

The purpose of a beaker to hold various types of

liquids. Such as in this experiment, this beaker
hold waste water.

2. 4 Bottles

3 bottles to be filled with certain amount of

waste water sample and will be fill full with
deionised water. I bottle will only fill with
deionised water as a blank sample.

3. Wastewater sample

The sample that will be use during experiment.

Source : G3 Lake, UTHM

4. Deionised Water

5. Pipettes

As a dropper
to transport a
measured
volume

of

liquid, often as
a media

dispenser.
To measured specific amount of wastewater.

6. pH Meter

A scientific
instrument that
measures the
hydrogenion
concentration
(pH) in a

solution
To measure the pH reading of wastewater sample

7. DO Meter

To measure dissolved oxygen (DO) in

a

8.

BOD Incubator
4

sample.

5.0
1.
2.

To keep the sample incubate at 20oC for 5 days

PROCEDURE
Waste water is collected from G3 Lake, UTHM.
The waste water is then tested by using pH meter. It is tested to be neutral. The

3.

readings for both pH and temperature are recorded.

The sample is then measured 2ml using pipette to pour in the bottle. 2 ml in each

4.

bottle for 3 bottles.

The sample is poured into the bottles first. Then pour the deionized water to the

5.
6.
7.
8.
9.

maximum of the bottles.

The forth bottle is prepared by pouring the deionized water only as blank sample.
DO meter is put into the bottles to get the DO reading for each bottle.
The readings of DO are recorded as initial DO.
The bottles are kept in the BOD incubator and remain around 20oc.
The final DO is recorded after 5 days of the experiment for every bottles.

10.

Determine the value of the BOD for each dilution using given formula.

6.0

RESULTS AND DATA ANALYSIS

Sample

:-

pH

: 7.1

Source

: Tasik G3

BOD Data Sample.

Date / Time Started : 09/03/16, 11AM

BIOCHEMICAL OXYGEN DEMAND ( BOD)

Analyst:

Date:02 / 03 / 16
Time : 11 A.M
Sample Details:
Source: Tasik Kemajuan (Tasik G3), UTHM
250C

7.1pH
Pretreatment:

Comments:

Alkalinity/Acidity

Neutral. This is because the sample water pH is 7.1 which is

Sample Volume:
I N NaoH :
I N N2SO4:

0
0

Sampl
e
Type

Sample
ID

BOD1

Blank

BOD2
BOD3

500

mL

considered as neutral. (6.5 7.5) = neutral

mL
mL
Volume
Sampl
e
( mL)

Dilution
Factor

Initial
DO
(mg/L)

Final
DO
( mg/L)

DO Depletion
( mg/L)

BOD
(mg/L)

0.83/207.
5 = 0.004

4.76

3.93

4.76 3.93
= 0.83

(0.83 x 500) = 207.5

2

Blank

0.004

4.98

4.25

0.73

182.5

Blank

0.004

5.56

5.10

0.46

115

Average BOD
( show the calculation)

Cancelled Data/ Result:

BOD1

= 207.5/3 = 69.17 mg/L

BOD2= 182.5/3 = 60.83 mg/L

BOD3= 115/3 = 38.33 mg/L

6.1

Calculate for the average BOD

( 207.5 + 182.5 + 115 ) = 168.33 mg/L

6.2

6.3

Show all the calculation and state if any of the data needs to throw out.

BOD1, DO depletion = DOi DOf = 4.76 3.93 = 0.83

BOD5 = ( 0.83 x 500 ) / 2 = 207.5 mg/L

BOD2, DO depletion = DOi DOf = 4.98 4.25 = 0.73 mg/L

BOD5 = ( 0.73 x 500 ) / 2 = 182.5 mg/L

BOD3, DO depletion = DOi DOf = 5.56 5.10 = 0.46 mg/L

BOD5 = ( 0.46 x 500 ) / 2 = 115 mg/L

The dilution water blank cannot deplete more than 0.2mg/L. Was this criteria
met ?
The BOD blank did not show a DO depletion of more than 0.2 mg/L after the five day
incubation period. A drop of more than 0.2 mg/L, indicates some type of
contamination or calibration error.

6.4

Does you sample shows a toxic effect ?

The sample is not showing any toxic effect, it is because the waste water, took from
Tasik G3 is not from a toxic source. The pH value of the sample is nearly neutral and
after our group use 5 mL of wastewater, the value of BOD obtained is slightly higher
than other groups. Its means that the sample doesnt show any toxic effect.

6.5

Could you rely on your BOD results? Why?

No. This is because some errors were occurred. Although the sample is occur from
toxicity, the dilution water blank does not meet the requirement. The dilution water
was prepared and stored for some times without proper quality control before the
experiment. This is caused growth of some biological that cannot be seen by naked
eyes. Existence of biological in the dilution water caused inaccuracy of the results.
Moreover, the wastewater used in BOD experiment does not undergo pre-treatment
process. It caused the wastewater dissolved heavy metal. The presence of heavy metal
make the result obtained may be different.

6.6

By referring to Sewage/ Effluent Standard (DOE Malaysia) could you

sample be discharge to river unrated? If not, suggest the associated
treatment for BOD removal.
According to the Sewage/ Effluent Standard (DOE Malaysia), sample cannot
discharge to the river unrated because the value obtained with 5ml of wastewater is
higher than 50 mg/L. the process which can used to treat wastewater in aeration.
Aeration is the process by which air is circulated through, mixed with or dissolved in
a liquid or substance. Normally, this treatment used in the secondary treatment of
wastewater. This may the higher dissolved oxygen content of wastewater will be
produced. Lastly, BOD level of wastewater will decrease.

7.0

DISCUSSION

QUESTION 1
a. Calculate the min / max volume of sample to be added to prepare as estimated
BOD :

400

mg/L

b. Why must samples that containing cautic alkalinity or acidity be adjusted before
preparing BOD dilution?
To conduct BOD experiment, the BOD dilution must be natural which range between
pH values of 6.5 to 7.0 because microbes are the most active in this range of pH value.
c. The complete composite sample arrive in the lab at (e.g). What is the latest day
and time the sample can be started for BOD?
The sample must be prepared not more than 24 hours. If it is begin within 2 hours of
collection, cold storage is unnecessary but, if the test will begin within more than 2 hours, the
sample must be keep at or below 4C during compositing.
d. Describe the function of BOD bottles cap and seal water.
BODs seal water was using to ensure that no air bubble is trapped in the bottle that
will affect the characteristics of sample. It is also to prevent further oxygen dissolving in.
Otherwise, bottles cap was using to reduce evaporate of the water seal during evaporation.

QUESTION 2
a. Why must samples that containing residual be dechlorinated before preparation
of BOD dilution?
The presence of chlorine in a sample will inhibit the growth of bacteria during the BOD test.

b. What reagents are required to chemically dechlorinated a BOD sample?

Sulphuric acid solution or acetic acid solution, potassium iodide solution, sodium sulphite
(Na2SO3).

c. What must be done to samples which have been dechlorinated or adjusted for
pH variations?
The samples which have been dechlorinated must be seeded with 300mL of BOD water in the
BOD bottle before allows it store in the BOD incubator.

QUESTION 3
a) State the formula to calculate
i)

Seed correction

= BOD5 of seed material x volume of sample / 300

ii)

BOD5 (seeded)

BOD5 (mg/L) = [(D1 D2) (B1 B2)] f] / P

Where ;
D1 = DO of diluted sample immediately after preparation, mg/L
D2 = DO of diluted sample after 5 days incubation at 20C, mg/L
P = decimal volumetric fraction of sample used
b) Calculate the seed correction and BOD5 (seeded) for the data given as below
BOD5 of Seed Material
mL of seed material

Dilution #1

95 mg/L
2 mL

mL of sample
100 mL
Start DO
7.8 mg/L
Final DO
2.9 mg/L
Seed correction = BOD5 of seed material x volume of sample / 300
=

95 x 2
mg
=0.633
<1.0 mg/ L
300
L

BOD5 mg/L = (D1 D2) / P

= [ ( 7.8 2.9 ) 0.633 mg/L ] / ( 100/300 )
= 12.801 mg/L
10

QUESTION 4
a) 30mL of wastewater are placed in a 300 mL BOD bottle. The sample is diluted to
fill the bottle. The DO concentrations at the beginning and the end of 5-day
incubation period are 7.3 mg/L and 1.8 mg/L respectively. What is the BOD ?
BOD5 = DOi - DOf / P = 7.3 1.8 / (30/300)
= 55 mg/L

b) The BOD5 of a wastewater was determined to be 250 mg/L. If the reaction

coefficient was 0.23 l/d, calculated
i)

Ultimate BOD

BODt = Lo (1 e-kt)
Lo = 250 / ( 1 e (0.23)(5))
= 365 mg/L
ii)

BOD3

BODt = Lo (1 e-kt)
BOD3 = 365( 1 e (0.23)(3))
iii)

= 181.9 mg/L

BOD remaining at 3 days

BOD8 = 365( 1 e (0.23)(3))

= 307 mg/L
QUESTION 5
a. What is seeding process in BOD measurement?
It is process of adding live bacteria and microorganism to a sample. If the samples
tested contain materials which could kill of injure the microorganism, the condition must be
corrected and healthy active organism added.

b. Explain properties of seed material.

11

Select a material to be used for seeding will have a BOD of at least 180 mg/L. This
will help to ensure that the seed correction meets the 0.6 mg/L minimum specified in
Standard Method.

Place the material in suitable container and incubator at 20C for 24-36 hours.
Usually, settled raw domestic sewage prepared in the manner above will have
sufficient BOD for use a seed material.

As an alternative, commercially available seed material may be used. The seed

correction should not exceed 1.0 mg/L BOD, therefore care should be taken not to use
too strong a seed material for the test.

The key to a good seed correction is a relatively stable seed material which produces
a good seed correction in every test situation.

c. What materials can be used to seed a BOD sample?

Any source of water which can provide a suitable population of organisms can be used,
however, settled raw sewage or commercially prepared seed material are the most common
sources.

QUESTION 6
What is the significance of dissolved oxygen?
Dissolved oxygen allows flora and fauna to breathe in water and it provides a suitable
habitat for the other animals. Bacteria in the water also use this oxygen to break down
animals and plants. The oxygen level is not very high in most water sources. If the oxygen
level is reduced, the animals begin to die and it is also to maintaining the aquatic life and
aesthetic quality streams and lakes.

12

QUESTION 7
a) With regard to precision, 10% duplicate or replicate samples should be run. This
would result in one duplicate sample or one replicate sample being run every ten
samples. Differentiate replicate and duplicate sample

Replicates have two or more separate water samples collected in the field from
the same site and depth. Replicate are used to determine the errors involved in
sample collection, if there are no errors in collection and analysis then
difference between two analysis indicates the natural variability in the water
at that location.

Duplicates have two or more lab analysis on the same water sample.
Duplicates are used to determine the % difference between two samples in
order to estimate the error involved in the analysis.

b) When are DO levels at their highest and their lowest and why?
DO levels are highest if the sample of water is not polluted means there is no toxic
and bacterial effect so in sample of water there is a lot of oxygen demanding wastes is well
within self-purification capacity. Besides the BOD will low if the water sample not have
microbes live and vice versa to the highest level of DO.

13

8.0

CONCLUSION
In conclusion, the objective of the experiment was achieved. The experiment was

carried out to measure the amount of dissolved oxygen in the water for a specified period of
the time and temperature. Normally BOD will be used to determine the BOD level in water.
However, in this experiment was used instead of BOD. By using BOD, the time of
experiment can be shorten and the effect to experiment almost the same with BOD. Since
average of BOD obtained from experiment is 168.33 mg/L, so that it can be concluded that
the wastewater from water plant at Tasik Kemajuan (G3), UTHM has been polluted. The
higher the BOD value, reading will result in lower DO level. When the DO level is lower, the
indicated that the water is unsuitable for living organisms. It must be treating for any possible
treatment to make sure that the water is safe and will not polluted by the pond.

14

9.0

REFERENCES
1. http://en.wikipedia.org/wiki/Dissolved_oxygen
2. http://www.ciese.org/curriculum/dipproj2/en/fieldbook/bod.shtml
3. http://www.apgqa.com/newsletters/6_2003_demand.asp
Jessica Raney, 2016.To Seed or Not To Seed - BOD Explained, Dilution, pH
Adjustment, Seeding and BOD Calculations.
4.

15

10.0

APPENDIX

16