ost Probable Number (
The most probable number (MPN) technique is an important technique in estimating microbial populations in soils, waters, and agricultural products. Many soils are heterogeneous, thereforeexact cell numbers of an individual organism can be impossible to determine. The MPNtechnique is used to estimate microbial population sizes in situations like this. The techniquedoes not rely on quantitative assessment of individual cells; instead it relies on specificqualitative attributes of the microorganism being counted. The important aspect of MPNmethodology is the ability to estimate a microbial population size based on a process-relatedattribute.This test method is a statistical, multi-step assay consisting of presumptive, confirmed andcompleted phases. In the assay, serial dilutions of a sample are inoculated into broth media.Analysts score the number of gas positive (fermentation of lactose) tubes, from which the other 2 phases of the assay are performed and then uses the combinations of positive results to consult astatistical tables to estimate the number of organisms present. Typically only the first 2 phasesare performed in coliform and fecal coliform analysis, while all 3 phases are done for E. coli.The 3-tube MPN test is used for testing most foods. The 5-tube MPN is used for water,
4.1. Presumptive coliform test
The presumptive coliform test is used to detect and estimate coliforms population of a water sample. In this test, a known volume of water is added to lactose fermentation tubes and production of acid and gas from the fermentation of lactose is a positive test for coliform bacteria. The lactose broth used in this test is selective for the isolation of coliforms. A pHindicator such as bromocresol purple is added to lactose broth for the detection of acid. Thecolour of pH indicator changes to yellow with the fermentation of acid from lactose.
Pond water sampleMcConkey broth of double and single with Durham tubes (5 tubes from double strength and 10from single strength)