ASTM D5465 - 16

    Standard Practices for Determining Microbial Colony Counts from Waters Analyzed by Plating Methods

    Active Standard ASTM D5465 | Developed by Subcommittee: D19.24

    Book of Standards Volume: 11.02

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    Significance and Use

    4.1 Numerous ASTM test methods and practices (for example: Test Methods D5259 and D5392, and Practices D6974 and E2563) report colony counts as their measured parameter.

    4.2 These practices provide a uniform set of counting, calculating, and reporting procedures for ASTM test methods in microbiology.



    A—Counting Colonies on Membrane Filters


    B—Counting Colonies on Pour Plates


    C—Counting Colonies on Spread Plates


    4.3 The counting rules provide a best attainable estimate of microorganisms in the sample, since the samples cannot be held and reanalyzed at a later date.

    1. Scope

    1.1 These practices cover recommended procedures for counting colonies and reporting colony-forming units (CFU) on membrane filters (MF) and standard pour and spread plates.

    1.2 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.

    1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

    2. Referenced Documents (purchase separately) The documents listed below are referenced within the subject standard but are not provided as part of the standard.

    ASTM Standards

    D1129 Terminology Relating to Water

    D5259 Test Method for Isolation and Enumeration of Enterococci from Water by the Membrane Filter Procedure

    D5392 Test Method for Isolation and Enumeration of Escherichia Coli in Water by the Two-Step Membrane Filter Procedure

    D6161 Terminology Used for Microfiltration, Ultrafiltration, Nanofiltration and Reverse Osmosis Membrane Processes

    D6974 Practice for Enumeration of Viable Bacteria and Fungi in Liquid Fuels--Filtration and Culture Procedures

    E2563 Practice for Enumeration of Non-Tuberculosis Mycobacteria in Aqueous Metalworking Fluids by Plate Count Method

    Other Standards

    9215 Heterotrophic Plate Count Available online from, or from American Public Health Association (APHA), Standard Methods for the Examination of Water and Wastewater, 800 I Street, NW Washington, DC 20001,

    ICS Code

    ICS Number Code 07.100.20 (Microbiology of water)

    Referencing This Standard
    Link Here
    Link to Active (This link will always route to the current Active version of the standard.)

    DOI: 10.1520/D5465-16

    Citation Format

    ASTM D5465-16, Standard Practices for Determining Microbial Colony Counts from Waters Analyzed by Plating Methods, ASTM International, West Conshohocken, PA, 2016,

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