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Membrane filter method for enumerating Escherichia coli

A P Dufour, E R Strickland, V J Cabelli

    Applied and Environmental Microbiology
    |May 1, 1981
    PubMed
    Summary
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    A new membrane filter method accurately detects Escherichia coli (E. coli) in water within 24 hours. This rapid technique eliminates the need for subculturing, improving efficiency in water quality testing.

    Area of Science:

    • Environmental microbiology
    • Water quality analysis
    • Bacteriology

    Background:

    • Accurate enumeration of Escherichia coli (E. coli) is crucial for assessing water safety.
    • Traditional methods for E. coli detection can be time-consuming, often requiring multiple steps like subculturing and biochemical identification.
    • There is a need for faster, more efficient methods to quantify E. coli in various water matrices.

    Purpose of the Study:

    • To develop and evaluate a rapid membrane filter procedure for the enumeration of Escherichia coli (E. coli).
    • To assess the efficiency and accuracy of the new method compared to existing techniques.
    • To provide a reliable method for E. coli quantification in marine, estuarine, and freshwater environments.

    Main Methods:

    • Development of a selective-differential medium for gram-negative, lactose-fermenting bacteria.

    Related Experiment Videos

  • Incorporation of a resuscitation step (2h at 35°C) prior to selective incubation (18-22h at 44.5°C).
  • Utilizing an in situ urease test for specific differentiation of E. coli from other thermotolerant, lactose-positive organisms.

    • Main Results:

    • The membrane filter procedure achieved E. coli recovery exceeding 90% across marine, estuarine, and freshwater samples.
    • Verification confirmed that 91% of presumptively positive colonies were indeed E. coli.
    • The method demonstrated high specificity, with less than 1% of verified E. coli colonies showing atypical reactions.

    Conclusions:

    • The developed membrane filter procedure offers a rapid and reliable method for enumerating E. coli in diverse water types.
    • This technique significantly reduces the time required for E. coli detection by eliminating the need for subculturing and isolate identification.
    • The high recovery and verification rates indicate the method's suitability for routine water quality monitoring and public health protection.