Theory

Biochemical oxygen demand, or BOD, measures the amount of oxygen consumed by

microorganisms in decomposing organic matter in stream water. BOD also measures the
chemical oxidation of inorganic matter (i.e., the extraction of oxygen from water via chemical
reaction). A test is used to measure the amount of oxygen consumed by these organisms
during a specified period of time (usually 5 days at 20 C). The rate of oxygen consumption in
a stream is affected by a number of variables: temperature, pH, the presence of certain kinds
of microorganisms, and the type of organic and inorganic material in the water.BOD directly
affects the amount of dissolved oxygen in rivers and streams. The greater the BOD, the more
rapidly oxygen is depleted in the stream. This means less oxygen is available to higher forms
of aquatic life. The consequences of high BOD are the same as those for low dissolved
oxygen: aquatic organisms become stressed, suffocate, and die.Sources of BOD include
leaves and woody debris; dead plants and animals; animal manure; effluents from pulp and
paper mills, wastewater treatment plants, feedlots, and food-processing plants; failing septic
systems; and urban stormwater runoff.

BOD is affected by the same factors that affect dissolved oxygen . Aeration of stream
water by rapids and waterfalls, for example will accelerate the decomposition of organic and
inorganic material. Therefore, BOD levels at a sampling site with slower, deeper waters
might be higher for a given volume of organic and inorganic material than the levels for a
similar site in highly aerated waters.Chlorine can also affect BOD measurement by inhibiting
or killing the microorganisms that decompose the organic and inorganic matter in a sample. If
you are sampling in chlorinated waters, such as those below the effluent from a sewage
treatment plant, it is necessary to neutralize the chlorine with sodium thiosulfate. BOD
measurement requires taking two samples at each site. One is tested immediately for
dissolved oxygen, and the second is incubated in the dark at 20 C for 5 days and then tested
for the amount of dissolved oxygen remaining. The difference in oxygen levels between the
first test and the second test, in milligrams per liter (mg/L), is the amount of BOD. This
represents the amount of oxygen consumed by microorganisms to break down the organic
matter present in the sample bottle during the incubation period. Because of the 5-day
incubation, the tests should be conducted in a laboratory.

Sometimes by the end of the 5-day incubation period the dissolved oxygen level is zero.
This is especially true for rivers and streams with a lot of organic pollution. Since it is not
known when the zero point was reached, it is not possible to tell what the BOD level is. In
this case it is necessary to dilute the original sample by a factor that results in a final
dissolved oxygen level of at least 2 mg/L. Special dilution water should be used for the
dilutions. It takes some experimentation to determine the appropriate dilution factor for a
particular sampling site. The final result is the difference in dissolved oxygen between the
first measurement and the second after multiplying the second result by the dilution factor.

(APHA. 1992. Standard methods for the examination of water and wastewater. 18th ed.
American Public Health Association, Washington, DC.)

The BOD5 test

Dilution method

To ensure that all other conditions are equal, a very small amount of micro-organism seed
is added to each sample being tested. This seed is typically generated by diluting activated
sludge with de-ionized water. The BOD test is carried out by diluting the sample with oxygen
saturated de-ionized water, inoculating it with a fixed aliquot of seed, measuring the
dissolved oxygen (DO) and then sealing the sample to prevent further oxygen dissolving in.
The sample is kept at 20 °C in the dark to prevent photosynthesis (and thereby the addition of
oxygen) for five days, and the dissolved oxygen is measured again. The difference between
the final DO and initial DO is the BOD. The apparent BOD for the control is subtracted from
the control result to provide the corrected value.The loss of dissolved oxygen in the sample,
once corrections have been made for the degree of dilution, is called the BOD5. For
measurement of carbonaceous BOD (cBOD), a nitrification inhibitor is added after the
dilution water has been added to the sample. The inhibitor hinders the oxidation of nitrogen.

BOD can be calculated by:

Undiluted: Initial DO - Final DO = BOD

Diluted: ((Initial DO - Final DO)- BOD of Seed) x Dilution Factor

BOD is similar in function to chemical oxygen demand (COD), in that both measure the
amount of organic compounds in water. However, COD is less specific, since it measures
everything that can be chemically oxidised, rather than just levels of biologically active
organic matter.
Manometric method

This method is limited to the measurement of the oxygen consumption due only to
carbonaceous oxidation. Ammonia oxidation is inhibited.The sample is kept in a sealed
container fitted with a pressure sensor. A substance that absorbs carbon dioxide (typically
lithium hydroxide) is added in the container above the sample level. The sample is stored in
conditions identical to the dilution method. Oxygen is consumed and, as ammonia oxidation
is inhibited, carbon dioxide is released. The total amount of gas, and thus the pressure,
decreases because carbon dioxide is absorbed. From the drop of pressure, the sensor
electronics computes and displays the consumed quantity of oxygen.

The main advantages of this method compared to the dilution method are:

simplicity: no dilution of sample required, no seeding, no blank sample,direct reading of

BOD value,continuous display of BOD value at the current incubation time.

Furthermore, as the BOD measurement can be monitored continuously, a graph of its

evolution can be plotted. Interpolation of several graphs on a similar water may build an
experience of its usual evolution, and allow an estimation of the five days BOD after as early
as the first two days of incubation .Most pristine rivers will have a 5-day carbonaceous BOD
below 1 mg/L. Moderately polluted rivers may have a BOD value in the range of 2 to 8 mg/L.
Municipal sewage that is efficiently treated by a three-stage process would have a value of
about 20 mg/L or less. Untreated sewage varies, but averages around 600 mg/L in Europe and
as low as 200 mg/L in the U.S., or where there is severe groundwater or surface water
infiltration
 Generally, when BOD levels are high, there is a decline in DO levels. This is because the
demand for oxygen by the bacteria is high and they are taking that oxygen from the oxygen
dissolved in the water. If there is no organic waste present in the water, there won't be as
many bacteria present to decompose it and thus the BOD will tend to be lower and the DO
level will tend to be higher.At high BOD levels, organisms such as macroinvertebrates that
are more tolerant of lower dissolved oxygen (i.e. leeches and sludge worms) may appear and
become numerous. Organisms that need higher oxygen levels (i.e. caddisfly larvae and
mayfly nymphs) will NOT survive.

(http://www.k12science.org/curriculum/dipproj2/en/fieldbook/bod.shtml)