FACULTY OF APPLIED SCIENCE

ENVIRONMENTAL SCIENCE & TECHNOLOGY

AS2293A
WATER RESOURCES TECHNOLOGY
EVT525
EXPERIMENT 2
BIOLOGICAL OXYGEN DEMAND (BOD)
(Respirometric Method)
NAME: FAIZAL BIN ALIAS
MATRIX NUMBER: 2014225942
DATE OF EXPERIMENT: 8th OCTOBER 2015
DATE OF SUBMISSION: 20rd OCTOBER 2015
NAME OF LECTURER: DR.NIK AZLIN NIK
ARIFIN

Introduction:
Biochemical Oxygen Demand is a common, environmental procedure for
determining the extent to which oxygen within a sample can support microbial
life. The following tutorial explores the theory and basics of performing this test
when one has little or no prior experience. This method is popular in many
environmental laboratories analyzing waste water, compost, sludge, and soil
samples. Although methods for each matrix are similar, this tutorial focuses on
the method associated with only waste water effluents.
The test for Biochemical Oxygen Demand is especially important in waste
water treatment, food manufacturing, and filtration facilities where the
concentration of oxygen is crucial to the overall process and end products. High
concentrations of dissolved oxygen (DO) predict that oxygen uptake by
microorganisms is low along with the required break down of nutrient sources in
the medium (sample). On the other hand, low DO readings signify high oxygen
demand from microorganisms, and can lead to possible sources of contamination
depending on the process.
Performing the test for Biochemical Oxygen Demand requires a significant
time commitment for preparation and analysis. The entire process requires five
days, and it is not until the last day where data is collected and evaluated.
During this time, samples are initially seeded with microorganisms and supplied
with a carbon nutrient source of glucose-glutamic acid. The sample is then
introduced to an environment suitable for bacterial growth at reproducible
temperatures, nutrient sources, and light within a 20 degree Celsius incubator
such that oxygen will be consumed. Quality controls, standards and dilutions are
also run to test for accuracy and precision. Determination of the dissolved
oxygen within the sample can be determined through Winkler titration methods.
The difference in initial DO readings (prior to incubation) and final DO readings
(after 5 days of incubation) predicts the BOD of the sample.

Objective:
To determine the value of BOD in water sample from three different places by
using respirometric method.

Procedure:
1) Three types of water sample are taken from three different places which is
section 6, section 7 and tap water in Shah Alam.
2) 6 BOD bottles are prepared and marked as blank, seed, standard, sample
1, sample 2 and sample 3.
3) Blank was added with 420ml BOD nutrients while seed was added with
355ml of seed and standard was added with 20ml of seed, 133ml of BOD
nutrients and 7ml of standard.
4) Sample 1, 2 and 3 were added with 35ml of seed and 320ml of samples
from three different places.
5) 3.8cm magnetic stir bar was placed in each sample bottles.
6) BOD nutrient buffer pillow was added to each bottle for optimum bacteria
growth.
7) Stopcock grease was applied to the seal lip and top of each seal for every
bottle.
8) Seal cup was placed in the neck of each bottle.
9) One lithium hydroxide powder pillow was added to the content of each
seal cup using a funnel.
10)
The bottles were placed on the base of the BOD Trak apparatus.
Appropriate tubes were connected to the sample bottle and firmly tighten
the cap.
11)
The instrument was placed into the incubator and instrument was
configured.
12)
The results are obtained and the data were tabulated after 6days.

BOD range (mg/l)

0-35
0-70
0-350
0-700

Required volume (ml)

420
355
160
95

Results and Data:

BOD bottles

1
2
3
4
5
6

Types

Blank
Seed
Standard
Sample 1
Sample 2
Sample 3

BOD bottles
Blank
Seed
Standard
Sample 1
Sample 2
Sample 3

Seed (ml)

355
20
35
35
35

BOD

Standard

Samples

Nutrients

(ml)

(ml)

(ml)
420
133
-

7
-

320
320
320

BOD Content (mg/L)

0.4
27.0
169.0
7.8
1.0
0.2

Location
Lake section 7
Lake at section 6
Tap water

Discussion:
Our group sample was from section 6 and the data reading by using YSI
for percentage of DO was 2.9, the temperature is 29.44C and pH is 8.62. So, our
samples was alkaline. The others water samples was taken from tap water, lake
at section 6 and lake at section 7.
For this experiment, the highest BOD content was in Standard which is
169.0 mg/L ad it was very polluted. The second highest was seed with BOD 27.0
mg/L. Then, the blank sample get lower BOD that is 0.4 mg/L and the water was
clean. Besides, there was the sample from another source which is Lake Section
7, Lake Section 6 and tap water. The highest BOD was sample 1 from Lake
Section 7 and this sample was not so polluted as in standard sample. The sample
3 has the lowest BOD with 0.2 mg/L it was clean. So, from these three samples,
the sample 3 was very suitable for use.
After that, our groups got the lower DO reading in sample 2, but after 5
days, our BOD reading was low. Its seems like the unbalance result was produced
in this experiment. So, maybe there was the error in this experiment that our

groups was done. This is because the lower DO must resulted in high BOD
reading logically.

Questions:
1. Why is it important to determine BOD value in wastewater treatment?

To prevent the threat of possible danger to health, or the very

existence of certain
species, it is essential to determine the
quality of a water source before water is drawn off for
consumption

2. Why do you incubate BOD bottles?

-

To exclude the light and prevent photosynthesis process happen.

3. Why do you need to determine the BOD of the blank (dilution water,
unseeded)?
-

It act as a control in the experiment.

4. What are the possible interferences in BOD determination? Explain how

you can overcome these interferences.
-

Incomplete carbon dioxide produced absorption that may affect cause

error if inappropriate amount of alkaline absorbent are not used.
Temperature fluctuation may also cause interferences in this
experiment.

5. Outline the principle of operation of the BOD Trak method.

-

the carbon dioxide is removed where pressure is proportional to the

amount of oxygen used.

the oxygen transferred to the bottle where the oxygen is transfer from

the air to the sample with during the stirring process.

The pressure sensor function helps to monitor air pressure within the
sample bottle. When the pressure begins to drop, the reading will
convert it to BOD reading.

6. Interpret your test results.

-

The BOD curve increase gradually as it gives a good results towards

each sample.

Conclusion:
It can be conclude that by using respirometric method, the BOD value for
blank, standard, seeded, sample 1, sample 2 and sample 3 solution was
determined after 5 days which is 0.4 mg/L, 27.0mg/L, 169.0mg/L, 7.8mg/L,
1.0mg/L and 0.2mg/L. This experiment was done successful but there were an
error done in the sample 2.

References:
Cairns, J. (1973). Biological methods for the assessment of water quality; a
symposium presented at the seventy-fifth annual meeting, American Society for
Testing and Materials. Philadelphia:

American Society for Testing and

Materials.
Langwith, J. (2010). Water. Detroit, MI: Greenhaven Press.
O'connor, R. (n.d.). Biological oxygen demand. Journal of Chemical Education,
209-209.
Muller, Mathieu; Bouguelia, Sihem; Goy, Romy-Alice; Yoris, Alison; Berlin, Jeanne;
Meche, Perrine; Rocher, Vincent; Mertens, Sharon; Dudal, Yves (2014).
"International cross-validation of a BOD5 surrogate". Environmental Science and
Pollution Research: 14. doi:10.1007/s11356-014-3202-3