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    Assignment On MPN

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    Noor-E-Khadiza Shama
    The document describes the most probable number (MPN) technique, a statistical method used to estimate microbes per unit volume in dilute samples like water. It involves using test tubes with samples at different dilutions, observing color change and gas production to detect growth. The result is interpreted using a reference table. The MPN test is completed in three steps - presumptive, confirmatory, and completed tests - to detect and confirm the presence of coliform bacteria like E. coli in water samples. The MPN technique is mostly used to test water quality and ensure its safety for consumption.

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    Assignment On MPN

    Uploaded by

    Noor-E-Khadiza Shama
    184 views3 pages
    The document describes the most probable number (MPN) technique, a statistical method used to estimate microbes per unit volume in dilute samples like water. It involves using test tubes with samples at different dilutions, observing color change and gas production to detect growth. The result is interpreted using a reference table. The MPN test is completed in three steps - presumptive, confirmatory, and completed tests - to detect and confirm the presence of coliform bacteria like E. coli in water samples. The MPN technique is mostly used to test water quality and ensure its safety for consumption.

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    Assignment on MPN

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    The document describes the most probable number (MPN) technique, a statistical method used to estimate microbes per unit volume in dilute samples like water. It involves using test tubes with samples at different dilutions, observing color change and gas production to detect growth. The result is interpreted using a reference table. The MPN test is completed in three steps - presumptive, confirmatory, and completed tests - to detect and confirm the presence of coliform bacteria like E. coli in water samples. The MPN technique is mostly used to test water quality and ensure its safety for consumption.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    184 views3 pages

    Assignment On MPN

    Uploaded by

    Noor-E-Khadiza Shama
    The document describes the most probable number (MPN) technique, a statistical method used to estimate microbes per unit volume in dilute samples like water. It involves using test tubes with samples at different dilutions, observing color change and gas production to detect growth. The result is interpreted using a reference table. The MPN test is completed in three steps - presumptive, confirmatory, and completed tests - to detect and confirm the presence of coliform bacteria like E. coli in water samples. The MPN technique is mostly used to test water quality and ensure its safety for consumption.

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    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    of 3

    Course: MCB302

    Section: 1
    Name: Noor-E-Khadiza Shama
    ID: 1921168
    Date: 08/30/20

    MPN Technique

    The most probable number (MPN) technique also known as multiple tube fermentation
    technique is a statistical method used to estimate the number of microbes per unit volume in
    dilute samples like water and food. It uses a specified number of test tubes with samples at
    different dilution factors, color change and gas production is observed at the end to detect
    growth. The result is interpreted by comparing with a reference table.

    MPN test is completed in 3 steps –

    1. 2. 3.
    Presumptive Confirmatory Completed
    Test Test Test

    1.Presumptive Test
    MPN usually utilizes 3 set method where each set refers to 5 test tubes. Out of 3 set, 2 set
    contains single strength liquid media (MacConkey lactose broth) and 1 set contains double
    strengthen liquid media (MacConkey lactose broth). Double strengthen refers to broth made up
    using twice the normal amount of broth powder. The following steps show how can we test a
    water sample for the presence of coliform bacteria -
     10 ml of double strengthen liquid media (MacConkey lactose broth) is placed in each
    test tubes of 1st set
     10 ml of single strengthen liquid media is placed in each test tubes of 2nd and 3rd set.
     Durham’s tube is inserted in inverted position in each test tubes of all sets.
     All the test tubes are then sterilized using autoclave for 15 minutes at 121 °
     After cooling water sample is added in each test tubes as follows-

    1. Add 10 ml water sample in each test tubes of 1st set


    2. Add 1 ml water sample in each test tubes of 2nd set
    3. Add 0.1ml water sample in each test tubes of 3rd

     Test tubes are incubate at 37°C for 24 hours.


     After incubation, the gas production is observed in Durham’s tube and color change of
    the media is also seen.
     The number of positive results from each set is recorded and compare with standard
    chart to give presumptive coliform count per 100 ml water sample.

    Reason for color change - The main selective factors found in the medium is lactose. Presence
    of coliform show color change because many bacteria cannot ferment this sugar, whereas
    coliform bacteria and several other bacterial types can ferment it.

    2. Confirmatory test
    Other microbes can also produce acid and gas from lactose fermentation, so to confirm the
    presence of coliform, confirmatory test is done. In this test, a small amount of suspension
    from positive tubes are inoculated into tubes with 3ml brilliant green lactose bile (BGLB).
    This is because BGLB inhibits growth of bacteria such as lactobacillus, Streptococci, Bacillus
    and Clostridium but allows coliforms to grow. The steps are as follows-

     One loopful of sample from each positive tubes is inoculated in respective tubes
    containing BGLB and incubated for 24 hours at 35.5 °
     Gas production 10% or more are recorded as positive while less than 10% is recorded
    as doubtful. Doubtful tubes are again incubated for 48 hours to check gas production.
    The results are recorded.
     At last, the number of coliform present in water sample is determined from previous
    MPN table.

    3. Completed Test
     It is a final test in which a loopful of sample from positive confirmatory tubes is streaked on
    Eosin methylene blue agar in a slant and incubated for 24 hours. The growth of gram-negative,
    non-sporing rods in the agar slants confirms the presence of coliforms.

    Significance of MPN
    It is mostly used in quality testing of water to ensure whether the water is safe for
    consumption. Though the presence of very few fecal coliform bacteria would indicate that
    consumption will not lead to any disease, but the presence of large numbers of fecal coliform
    bacteria would indicate that it is unsafe for consumption. It is also used in estimating microbial
    populations in agricultural products. Through the MPN method, turbid water containing
    sediments, sludge, mud etc can be analysed.

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