OBJECTIVE

For biochemical oxygen demand (BOD) experiment, the objective is to

measure the amount of dissolved oxygen in the wastewater for a specific
period of time and temperature.

INTRODUCTION
Biochemical Oxygen Demand is a common, environmental procedure for
determining the extent to which oxygen within a sample can support microbial
life. The microorganisms use organic matter as a food source through oxidation
which oxygen is consumed. According to Linda (1994), 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. . The study orientation was on
the beneficial uses of water which was focused on, water for domestic water
supply, fisheries and aquatic propagation, livestock drinking, recreation and
agricultural use (DOE Malaysia, 1985).
The higher the BOD, the more oxygen will be demanded from the waste to
break down the organics. The BOD test is performed by incubating a sealed
wastewater sample for the standard 5-day period, then determining the change
in dissolved oxygen content. The bottle size, incubation temperature, and
incubation period are all specified. Most wastewaters contain more oxygen
demanding materials than the amount of DO available in air-saturated water.
Therefore, it is necessary to dilute the sample before incubation to bring the
oxygen demand and supply into appropriate balance. Because bacterial growth
requires nutrients such as nitrogen, phosphorous, and trace metals, these are
added to the dilution water, which is buffered to ensure that the pH of the
incubated sample remains in a range suitable for bacterial growth. Complete
stabilization of a sample may require a period of incubation too long for practical
purposes, therefore, 5-day period has been accepted as the standard incubation
period. Quality controls, standards and dilutions are also run to test for accuracy
and precision (Linda, 1994).
There are some the environmental impacts of BOD which is the sewage
containing high BOD create environmental and health problems. Moreover, it
interferes with the aquatic life. Organic pollution is harmful to fish as it tends to
reduce the amount of dissolved oxygen. Last but not least, it defines the strength
of domestic wastes and industrial wastewaters
APPARATUS & MATERIALS

6 units of 300mL BOD bottles with glass stoppers

Volumetric pipette
Incubator at 25C

REAGENTS

Dilution water containing nutrient buffer and seed

Water sample

PROCEDURE
1. The dilution water was prepared using a BOD nutrient buffer pillow.
2. The experiment was divided into seeded and unseeded dilution
water.
3. For seeded dilution water, the BOD bottle was filled up with the
dilution water and was labeled as blank seeded sample. For other
two BOD bottles were filled up with dilution water as well as
inserted the 3mL and 6mL of water sample to each of the BOD
bottles.
4. For each of BOD bottles were added nutrients and seed.
5. Step 3 was followed for unseeded dilution water.
6. The bottles were closed with stopper. The stopper was tightly
twisted into place and pressed down and inverted the bottles
several times to mix.
7. The initial dissolved oxygen concentration was measured in each of
bottle both for unseeded and seeded sample.
8. The dilution water was added to the lip of the BOD bottles to make
water seal.
9. The bottles were incubated in the incubator for 7 days at 20C.
10.
After 7 days, the remaining dissolved oxygen concentration
was measured in each bottle.
11.
The BOD value was calculated.
RESULT
To calculate the BOD5 value,
For unseeded dilution water

BOD 5=

( DO initial DO final ) bottle volume

s ample volume

For seeded dilution water

BOD 5=

( D 1D 2 ) (B 1B2) f
P

Where;
D1=initial DO concentration of sample, mg/L
D2= final DO concentration of sample, mg/L

B1= initial DO concentration of blank, mg/L

B2= final DO concentration of blank, mg/L
f= fraction seeded dilution water volume in sample to blank
P= decimal fraction of sample in 300mL bottle

Sample,
mL
Seeded

Blank

DO
initial,
mg/L
7.28

DO
final,
mg/L
7.15

BOD7, mg/L

300
3000/

mL
mg
mg
7.497.23
(7.287.15
)
L
L

0
3mL
sample

7.33

7.22

3003

300

mg
mg
7.337.22
(7.287.15
)
L
L

1.87
6mL
sample

7.28

7.37

300
3006/

300 mL

300
6/

mg
mg
7.287.37
(7.287.15
)
L
L

10.87
Unseede
d

Blank

7.49

7.23

mg
) 300 mL
L
0 mL

(7.497.23

3mL
sample

7.34

mg
)300 mL
L
3 mL

7.09

(7.347.09

25 .0
6mL
sample

7.33

mg
)300 mL
L
6 mL

7.06

(7.337.06

13.5

Calculation for BOD5

The formulae to calculate BOD5 is;

Ultimate BOD=BOD 5/ ( 1e(k ) (T ) )

BOD 5=Ultimate BOD ( 1e( k ) (T ) ) where
k= reaction constant per day
T= time

Unseede
d

Sample,
mL
Blank

BOD7,
mg/L
0

Ultimate BOD, mg/L

0/(1e(0.23) (7 ))
0

3mL
sample

25

13.5

25.0/(1e(0.23) (7 ))

Blank

13.5/(1e( 0.23) (7 ))

e(0.23)(5)
16.875
11.53

( 0.23) ( 7 )

0/(1e
=0

1e(0.23 )(5)
31.25
21.36

16.875
Seeded

1e(0.23 )(5)
0
0

31.25
6mL
sample

BOD5, mg/L

1e(0.23 )(5)
0
0

3mL
sample

-1.87

1.87 /(1e( 0.23) (7 ))

2.34

6mL
sample

-10.87

10.87 /(1e( 0.23) (7 ))

13.6

1e(0.23 )(5)
2.34
1.60

1e(0.23 )(5)
13.6
9.29

DISCUSSION
On the BOD experiment, we have gathering the BOD results. Theoretically,
the BOD value is calculate in 5 days but in this experiment the BOD value is
indicated until day 7. So, we are using the ultimate BOD formulae to find the
exact value of BOD5. According to INQWS (Index National Quality Water
Standards), the good BOD5 value is less than 5ppm, which is suitable for human
daily consumption. 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 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.
The result showed that for unseeded sample, is in higher numbers,
indicate that the water sample is highly contaminated. A high BOD indicates a
high content of easily degradable, organic material in the sample. Apparently,
the sample water for 3mL is in Class V since the BOD 5 value is more than
12ppm,whereas for 6mL sample water is in Class IV, based on DOE Water Quality
Index Classification. This showed the dissolved oxygen is depleted since there
are many microbiological organisms that have in the water sample. Thus, it can
be harmful to human and environment condition.
Theoretically, for BOD5, the number of seeded must be higher than seeded
because in seed packet contain nutrient consists of phosphorus and nitrogen
which is the major metal for growth of microbiology organisms. Hence, it should
consume a lot of oxygen than unseeded sample. But in this case, the value for
seeded sample for both 3mL and 6mL of water sample showed the negative
results. This may due to some errors during the lab experiment. The problem
may come from diluting the BOD sample. During the time to fill up the BOD
bottles, the student should take turns interspersed the sample and dilution
water, by the same time shake the bottle regularly in order for the mixture to be
mix together before taking the DO value.

CONCLUSION

From the data collected above, the value of BOD 5 for unseeded 3mL and 6mL of
water sample are 21.36mg/L and 11.53mg/L respectively. For seeded 3mL and
6mL of water samples are -1.60mg/L and -9.29mg/L respectively.
Therefore, as the conclusion, only value of BOD5 for unseeded 3mL and 6mL of
water sample has polluted and contaminated sample, whereas for seeded
showed an error during the experiment.

REFERENCES
Department of Environment Malaysia, (1985). Development of Water Quality and
Standards for Malaysia
Linda S. M. (1994). Biochemical Oxygen Demand. Retrieved from
http://www.rpi.edu/dept/chem-eng/Biotech-Environ/Environmental/BOD/coda.htm