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Spring 2019
Date: 4/2/2019
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INTRODUCTION
The objective of this lab is to learn how to conduct BOD5 tests. Biochemical Oxygen
Demand (BOD) i s t h e a m o u n t o f o x y g e n a b a c t e r i u m n e e d s t o a e r o b i c a l l y
o x i d i z e o r g a n i c w a s t e s . B O D i s a m o n g t h e m e t h o d s u s e d in quantifying water
pollution. W h e n o rganic wastes are disposed into water, they may either be suspended or totally
dissolved (Jing L. et. Al, 2002). Presence of oxygen and organic matter facilitate the live and
continuous growth of bacteria in the water because of provision of oxidation and food respectively.
Animals and bacteria oxidized organic matter by breaking them down with existence of oxygen
in their systems either by digestion or biochemical action of bacteria. Usually, to determine the
water sample BOD, a calculation of the amount of oxygen used by the bacteria is performed by
comparing the amount left at the final fifth day with the initial amount at the first day. At the end
of the fifth day, bacteria will have oxidized mostly the soluble organic matter available while a
very slight oxidation of the insoluble matter will take place (Michael R. et al, 2009).
The degree of water pollution is usually determined basically means BOD measurements.
This is the most vital measurement taken in managing a sewage treatment plant. Essentially, the
effectiveness and efficiency of sewage treatment can be studied by making a comparison of the
BOD of sewage into the plant and the BOD of treated water leaving the plant. If water with a high
content of BOD gets into a river, it will enable the bacteria in the river to oxidize the organic
matter, the oxygen from the river will be consumed faster than the rate it dissolves back in from the
river causing aquatic life to die out due to lack of oxygen (Jun H. et al, 2000). It is essential for
sewage treatment plants eliminate as much BOD as possible from the sewage water.
𝑆𝑎𝑚𝑝𝑙𝑒 𝑉𝑜𝑙𝑢𝑚𝑒(𝑚𝐿)
𝐷𝑖𝑙𝑢𝑡𝑖𝑜𝑛 𝑓𝑟𝑎𝑐𝑡𝑖𝑜𝑛 =
𝐵𝑜𝑡𝑡𝑙𝑒𝑑 𝑣𝑜𝑙𝑢𝑚𝑒(300 𝑚𝐿)
METHODS
The materials used in this lab were; 300mL BOD bottles, large plastic bottles with siphons, DO meter
and graduated cylinders. The prepared nutrients solutions provided include; Phosphate buffer, Magnesium
sulfate solution, Calcium chloride solution, Ferric chloride solution and Ammonium Chloride solution.
Firstly, the desired volume of DI water was added to a 20-L on a plastic 20 L carboy. Water was
aerated overnight by bubbling in ambient air compressed air. The saturated O2 concentration in the water
was around 8.4 mg/L DO. 1 mL per liter of the five nutrients solutions were added to the aerated water.
The appropriate volume of well mixed diluted sample was transferred into a well cleaned and rinsed
300mL BOD bottles using a tipped, volumetric pipette. BOD was slowly filled ensuring there was no air
bubble trapped inside. Triplicate bottles were also filled with dilution water. The bottles were carefully
labelled. The calibrated DO meter was used to measure the initial DO of the dilution water blanks and the
measure recorded. BOD bottles were placed in a cabinet at room temperature for 5 days. The calibrated DO
meter was again used to measure the final DO of the dilution water blanks and the sample bottles and the
measure recorded.
.
RESULTS
For the determination of biochemical oxygen demand (BOD) of wastewater, the BOD is
computed subtracting the final DO from the initial DO and dividing by the dilution fraction. The
test measures the ability of bacteria to digest organic matter after five days at room temperature
𝑆𝑎𝑚𝑝𝑙𝑒 𝑉𝑜𝑙𝑢𝑚𝑒(𝑚𝐿)
𝐷𝑖𝑙𝑢𝑡𝑖𝑜𝑛 𝑓𝑟𝑎𝑐𝑡𝑖𝑜𝑛 =
𝐵𝑜𝑡𝑡𝑙𝑒𝑑 𝑣𝑜𝑙𝑢𝑚𝑒(300 𝑚𝐿)
These formulas were used in excel to calculate BOD5 and the following results were obtained.
Initial DO Final DO dilution BOD5
(mg/L) (mg/L) fraction (mg/L)
2.6 0 390
2.4 0 360
volume 2mL 0.006666667
1.9 0 285
2 0 300
7.6 5.7 190
9.4 6 340
volume 3mL 0.01
7.6 5 260
8.6 4.9 370
8.2 4.7 262.5
8.8 4.3 337.5
volume 4mL 0.013333333
8.2 4.7 262.5
8.6 5 270
8.1 2.9 312
7.9 2.3 336
volume 5mL 0.016666667
8.1 2.7 324
7.9 3 294
7 3.7 132
volume 6.8 0.5 252
0.025
7.5mL 6.7 2.5 168
250
200
150
100
50
0
0 2 4 6 8 10 12 14
Sample water volume (mL)
need an environment that is suitable to survival. The dissolved oxygen in all the samples
decreased at the end of the five days. It is observed that the concentration of dissolved oxygen
decreases a s the dilution of the samples increased. This is because there is a high demand of
oxygen from bacteria. After fifth day, the dissolved oxygen became less as compared to the first
day. A high d e m a n d of oxygen from bacteria can be proven with dissolved oxygen
contamination sources.
Table 2. From the graph in Fig 1, it is observed that the BOD for the samples generally reduced
with the increasing dilution of volume of the samples. The lesser the BOD value, the lower the
sample is polluted. High amount of BOD shows a high amount of organic material in the sample
that is easily degradable. While, a low BOD shows less amount of organic materials matter, that
are difficult to dissolve. The dilutions according to the BOD5 range provided in the lab manual
indicate the results obtained in this experiment are realistic since the fall in that range.
The quality of wastewater is very polluted when the BOD5 level is greater than 100, while
the wastewater quality is somewhat polluted when the BOD5 level is between 6 to 9. Hence, the
BOD5 level of all dilutions were above 100 ppm showing that all the dilution was very polluted.
Nitrification is the by which specific bacteria oxidize nitrogen from ammonium to nitrate.
The nitrification inhibitor prevents the specific bacteria that are accountable for the deprivation of
the ammonium without damaging the microorganisms that degrade carbon compounds.
ammonium to nitrate. In this experimental, the objective of learning how to conduct BOD5 tests
was successfully achieved. BOD of all wastewater samples were all determined.
REFERENCES
Jing Liu and Bo Mattiasson. (2002). Microbial BOD Sensors for Wastewater Analysis. Journal of
Michael R. Penn, James J. Pauer and James R. Mihelcic. (2009). Biochemical Oxygen Demand.
Jun Ho. Lee and Ki Woong Bang. (2000). Characterization of Urban Stormwater Runoff. Journal
Initial DO Final DO
(mg/L) (mg/L)
2.6 0
2.4 0
volume 2mL
1.9 0
2 0
7.6 5.7
9.4 6
volume 3mL
7.6 5
8.6 4.9
8.2 4.7
8 4.3
volume 3mL
8.2 4.7
8.6 5
8.1 2.9
7.9 2.3
volume 5mL
8.1 2.7
7.9 3
7 3.7
volume 6.8 0.5
7.5mL 6.7 2.5
7.8 1.1
volume 8.4 1.2
10mL 8.2 1
8.3 1
10 0.1
volume 10.1 0.2
12.5mL 9.9 0.1
10 0
𝑆𝑎𝑚𝑝𝑙𝑒 𝑉𝑜𝑙𝑢𝑚𝑒(𝑚𝐿) 2 𝑚𝐿
𝐷𝑖𝑙𝑢𝑡𝑖𝑜𝑛 𝑓𝑟𝑎𝑐𝑡𝑖𝑜𝑛 = = = 0.006666667
𝐵𝑜𝑡𝑡𝑙𝑒𝑑 𝑣𝑜𝑙𝑢𝑚𝑒(300 𝑚𝐿) 300 𝑚𝐿