MEASURING BIOLOGICAL OXYGEN DEMAND (BOD)

Very little oxygen dissolves in water and its solubility actually decreases with increasing
temperature. If water becomes polluted with organic waste, such as raw sewage, farm slurry or silage
effluent, or detergents containing nitrates and phosphates, its oxygen content will quickly be
depleted, a condition known as eutrophication. Eutrophication occurs because the organic wastes
enable the rapid growth of algae and phytoplankton. When these photosynthesisers die, the aerobic
bacteria present in the water oxidise and decompose the organic matter and dead plants, so removing
oxygen from the water and creating a body of water in which larger organisms cannot survive.

In warm weather, all the oxygen dissolved in a river or stream at a point of heavy pollution, may be
used up, causing the death of all aerobic, aquatic organisms, including the fish, for some distance
downstream.

The BIOLOGICAL OXYGEN DEMAND (BOD) is the amount of oxygen (mg dm-3) consumed in
a sample of water kept in the dark for 5 days at 20°c.

BOD is formally calculated by collecting two samples of water at the same time from the same
source. One sample is tested immediately to determine the dissolved oxygen concentration, The
other sample is left in the dark at 20°c for 5 days, and then tested for dissolved oxygen concentration.
The BOD is the difference between the dissolved oxygen concentration in the two samples.

Determining the BOD in this way may not be feasible in a school lab. Either specific probes are
required or a technique known as the Winkler Method must be employed, using various buffered
solutions. An alternative and simple method to compare the BOD of two water samples uses
methylene blue dye, which goes colourless as soon as oxygen is absent from a sample of water. In
this way, it is possible therefore to compare the BOD of water upstream and downstream of a point
source of pollution.

LAB
Aim
To compare the Biological Oxygen Demand (BOD) of two samples of water, upstream and
downstream of point source of water pollution.

Risk assessment
This lab has few risks, except perhaps in the collection of water samples from a river or lake, where
care is necessary.

Research
See if it possible to determine the type of the organic pollution in the water you are sampling. The
pollution may come from fertilisers or farm slurry or detergents or sewage, or from some sort of
manufacturing process.

Hypothesis
Make a hypothesis, comparing the two water samples you have collected. Justify the hypothesis by
referring to the process of eutrophication and its effects on water at a point of pollution.
Equipment and materials
 Two 250ml glass bottles with ground-glass stoppers, suitable for collecting air-free samples.
 1 ml pipette or syringe
 Solution of 0.1% methylene blue
 Temperature controlled (20°c), dark environment to store samples

Method
1. Collect the two samples of water in 250ml bottles with ground-glass stoppers. The samples
should be collected from the same depth and in a way that avoids agitation of the water.
When the bottles are stoppered, there should be no air present.
2. Add 1ml of 0.1% methylene blue solution to each sample by using a pipette or syringe that
reaches to the bottom of the bottles. Replace the stoppers, ensuring no air is trapped in the
bottles.
3. Place the bottles together in a dark environment at 20°c.
4. Observe the bottles every day, if possible morning and evening, and record the time taken
for each bottle to go colourless.

Data presentation
Present the results as a simple bar graph.

Analysis and conclusion

The determination of the BOD may be affected by other pollutants such as heavy metals, which can
inhibit bacterial metabolism, or by the specific sort of organic pollutants. Nevertheless, you should
be able to make an analysis of your preliminary research, the results and the graphs, sufficient to
state a conclusion.

http://slideplayer.com/slide/4629580/

John Osborne
March 2018