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Detection and Enumeration of the MPN

Detection and Enumeration of the MPN

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Published by: kolita kamal on Aug 13, 2010
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DETECTION ANDENUMERATION OF THE MOSTPROBABLE NUMBER (MPN) OFCOLIFORM BACTERIA IN ASAMPLE OF WATER
 
GS/M.Sc./FOOD/3608/08
 
B.K.K.K.Jinadasa
 
 
 
1
Introduction
Coliforms are normally present on raw produce, and their presence does not necessarily signifythat pathogens are present or that the produce has come in contact with feces.The coliform group includes species from the genera Citrobacter, Klebsiella, Enterobacter, andE.coli. Other than E. coli, no foodborne outbreaks have been associated with coliforms.Coliforms were historically used as "indicator microorganisms" to serve as a measure of fecalcontamination, and thus potentially of the presence of enteric pathogens. Although somecoliforms are found in the intestinal tract of man, most are found throughout the environment andhave little sanitary significance. The presence of indicator organisms such as E. coli andcoliforms can be an indication of faecal contamination arising from within the slaughter facilityor post slaughter, inadequate processing, post-processing contamination or a combination of anyor all of the above.Coliform counts are inadequate to differentiate between faecal and non-faecal contamination. E.coli is considered to be more closely associated with faecal contamination from warm- bloodedvertebrates than are other members of the coliforms. Hence, the presence of E. coli in food or water became accepted as indicative of recent faecal contamination and the possible presence of frank pathogens.In an effort to indicate µquality¶ or µsafety¶ of such things as dairy products, fruits, vegetables,drinking water, composted manure, and treated sewage, microbiologists developed the positiveassociation of groups of bacteria to fecal contamination and dubbed this group of bacteria ''faecalcoliform" . The term "thermotolerant coliforms" has recently been used to describe theseorganisms and is more appropriate. These general terms for a large and diverse class of bacteriaare useful and remain relevant in specific food, wastewater management, and water qualityapplications. The ability to rapidly and accurately detect E. coli and coliform bacteria isimportant to any food quality and safety program.
 
 
2
M
ost Probable Number (
M
PN)
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.
M
aterials
Pond water sampleMcConkey broth of double and single with Durham tubes (5 tubes from double strength and 10from single strength)

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