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Biochemical oxygen demand or BOD is a chemical procedure for determining the uptake rate of dissolved oxygen by the biological organisms in a body of water. It is not a precise quantitative test, although it is widely used as an indication of the quality of water.

BOD can be used as a gauge of the effectiveness of wastewater treatment plants. It is listed as a conventional pollutant in the U.S. Clean Water Act.

The BOD5 test

BOD measures the rate of oxygen uptake by micro-organisms in a sample of water at a temperature of 20°C and over an elapsed period of five days in the dark.

There are two recognized methods for the measurement of BOD.

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.

Test Limitations

The test method involves variables limiting reproducibility. Tests normally show observations varying plus or minus ten to twenty percent around the mean.


Some wastes contain chemicals capable of suppressing microbial growth or activity. Potential sources include industrial wastes, antibiotics in pharmaceutical or medical wastes, sanitizers in food processing or commercial cleaning facilities, chlorination disinfection used following conventional sewage treatment, and odor-control formulations used in sanitary waste holding tanks in passenger vehicles or portable toilets. Suppression of the microbial community oxidizing the waste will lower the test result.

Appropriate Microbial Population

The test relies upon a microbial ecosystem with enzymes capable of oxidizing the available organic material. Some waste waters, such as those from biological secondary sewage treatment, will already contain a large population of microorganisms acclimated to the water being tested. An appreciable portion of the waste may be utilized during the holding period prior to commencement of the test procedure. On the other hand, organic wastes from industrial sources may require specialized enzymes. Microbial populations from standard seed sources may take some time to produce those enzymes. A specialized seed culture may be appropriate to reflect conditions of an evolved ecosystem in the receiving waters.

History of the use of BOD

The Royal Commission on River Pollution, which was established in 1865 and the formation of the Royal Commission on Sewage Disposal in 1898 led to the selection in 1908 of BOD5 as the definitive test for organic pollution of rivers. Five days was chosen as an appropriate test period because this is supposedly the longest time that river water takes to travel from source to estuary in the U.K.marker In 1912, the commission also set a standard of 20 ppm BOD5 as the maximum concentration permitted in sewage works discharging to rivers, provided that there was at least an 8:1 dilution available at dry weather flow. This was contained in the famous 20:30 (BOD:Suspended Solids) + full nitrification standard which was used as a yardstick in the U.K. up to the 1970s for sewage works effluent quality. The value of BOD is less then COD.

The United Statesmarker includes BOD effluent limitations in its secondary treatment regulations. Secondary sewage treatment is generally expected to remove 85 percent of the BOD measured in sewage and produce effluent BOD concentrations with a 30-day average of less than 30 mg/L and a 7-day average of less than 45 mg/L. The regulations also describe "treatment equivalent to secondary treatment" as removing 65 percent of the BOD and producing effluent BOD concentrations with a 30-day average less than 45 mg/L and a 7-day average less than 65 mg/L.

Typical BOD values

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.marker, or where there is severe groundwater or surface water infiltration. (The generally lower values in the U.S. derive from the much greater water use per capita than in other parts of the world.)



  1. U.S. Environmental Protection Agency (EPA). Washington, DC. "Secondary Treatment Regulation." Code of Federal Regulations, 40 CFR Part 133.

See also

External links

  • BOD Doctor - a troubleshooting wiki for this problematic test

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