Sampling Considerations

BOD is affected by the same factors that affect dissolved oxygen (see above). 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. (See APHA, 1992.) 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. (See APHA, 1992.) 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. More details are provided in the following section.

As BOD5 depends on the growth of microbes the test can be adversely affected by

Temperature Toxics Turbidity Light

Agitation Species of Microbe Microbial contamination by a wild population Nature of the organics present Skill of the analyst Data entry errors Lack of blanks and samples of known strength Equipment accuracy and maintenance