ASTM D5392-24
(Test Method)Standard Test Method for Isolation and Enumeration of Escherichia coli in Water by the Two-Step Membrane Filter Procedure
Standard Test Method for Isolation and Enumeration of <emph type="bdit">Escherichia coli</emph> in Water by the Two-Step Membrane Filter Procedure
SIGNIFICANCE AND USE
5.1 This test method is useful for measuring recreational water quality and chlorinated wastewaters, although it can be used for any water suspected of contamination by fecal wastes of warm-blooded animals. The significance of finding E. coli in recreational water samples, especially samples obtained from fresh recreational waters, is that there is a risk of gastrointestinal illness, directly related to the E. coli density, associated with swimming.5
5.2 Since small or large volumes of water or dilutions thereof can be analyzed by the MF technique, a wider range of levels of E. coli in water can be detected and enumerated than with other methods.
SCOPE
1.1 This test method describes a membrane filter (MF) procedure for the detection and enumeration of Escherichia coli, a bacterium found exclusively in the feces of humans and other warm-blooded animals. The presence of these microorganisms in water is an indication of fecal pollution and the possible presence of enteric pathogens. These bacteria are found in water and wastewater in a wide range of densities. The detection limit of this procedure is one colony forming unit (CFU) per volume filtered.
1.2 This test method has been used successfully with temperate fresh and marine ambient waters, and wastewaters. It is the user’s responsibility to ensure the validity of this test method for waters of other types.
1.3 The values stated in SI units are to be regarded as standard.
1.4 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, health, and environmental practices and determine the applicability of regulatory limitations prior to use. For specific hazard statements, see Section 9.
1.5 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
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This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation: D5392 − 24
Standard Test Method for
Isolation and Enumeration of Escherichia coli in Water by
1
the Two-Step Membrane Filter Procedure
This standard is issued under the fixed designation D5392; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope D1193 Specification for Reagent Water
D3370 Practices for Sampling Water from Flowing Process
1.1 This test method describes a membrane filter (MF)
Streams
procedure for the detection and enumeration of Escherichia
D3870 Practice for Establishing Performance Characteristics
coli, a bacterium found exclusively in the feces of humans and
for Colony Counting Methods in Microbiology (With-
other warm-blooded animals. The presence of these microor-
3
drawn 2000)
ganisms in water is an indication of fecal pollution and the
D5465 Practices for Determining Microbial Colony Counts
possible presence of enteric pathogens. These bacteria are
from Waters Analyzed by Plating Methods
found in water and wastewater in a wide range of densities. The
detection limit of this procedure is one colony forming unit
3. Terminology
(CFU) per volume filtered.
3.1 Definitions:
1.2 This test method has been used successfully with
3.1.1 For definitions of terms used in this standard, refer to
temperate fresh and marine ambient waters, and wastewaters. It
Terminology D1129.
is the user’s responsibility to ensure the validity of this test
method for waters of other types.
3.2 Definitions of Terms Specific to This Standard:
3.2.1 Escherichia coli (E. coli), n—a species of bacteria that
1.3 The values stated in SI units are to be regarded as
is a member of the total coliform group and known to originate
standard.
in the feces of warm-blooded animals.
1.4 This standard does not purport to address all of the
safety concerns, if any, associated with its use. It is the 3.3 Performance Characteristics (Practice D3870):
responsibility of the user of this standard to establish appro-
3.3.1 accuracy, n—the proportion of the observed count to
priate safety, health, and environmental practices and deter-
the true density of a sample.
mine the applicability of regulatory limitations prior to use.
3.3.2 bias, n—the persistent positive or negative deviation
For specific hazard statements, see Section 9.
of the average value of the test method from the assumed or
1.5 This international standard was developed in accor-
accepted true value.
dance with internationally recognized principles on standard-
3.3.3 precision, n—the degree of agreement of repeated
ization established in the Decision on Principles for the
measurements of the same parameter expressed quantitatively
Development of International Standards, Guides and Recom-
as the standard deviation or as the 95 % confidence limits of
mendations issued by the World Trade Organization Technical
the mean computed from the results of a series of controlled
Barriers to Trade (TBT) Committee.
determinations.
2. Referenced Documents
3.3.4 specificity, n—the ability of a test method to select or
2
distinguish, or both, the target bacteria in the same water
2.1 ASTM Standards:
sample; the specificity characteristic of the method is usually
D1129 Terminology Relating to Water
reported as the percent of false positive and false negative
results.
1
This test method is under the jurisdiction of ASTM Committee D19 on Water
3.3.5 upper counting limit (UCL), n—that colony count
and is the direct responsibility of Subcommittee D19.24 on Water Microbiology.
above which there is an unacceptable counting error; the error
Current edition approved April 1, 2024. Published April 2024. Originally
may be due to overcrowding or antibiosis.
approved in 1993. Last previous edition approved in 2019 as D5392 – 19. DOI:
10.1520/D5392-24.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
3
Standards volume information, refer to the standard’s Document Summary page on The last approved version of this historical standard is referenced on
the ASTM website. www.astm.org.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 ----------------------
D5392 − 24
4. Summary of Test Method 7.2 Microscope Lamp, producing diffuse light from a cool,
4 white fluorescent lamp adjusted to give maximum visibility.
4.1 This two-step test method p
...
This document is not an ASTM standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Because
it may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current version
of the standard as published by ASTM is to be considered the official document.
Designation: D5392 − 19 D5392 − 24
Standard Test Method for
Isolation and Enumeration of Escherichia coli in Water by
1
the Two-Step Membrane Filter Procedure
This standard is issued under the fixed designation D5392; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope
1.1 This test method describes a membrane filter (MF) procedure for the detection and enumeration of Escherichia coli, a
bacterium found exclusively in the feces of humans and other warm-blooded animals. The presence of these microorganisms in
water is an indication of fecal pollution and the possible presence of enteric pathogens. These bacteria are found in water and
wastewater in a wide range of densities. The detection limit of this procedure is one colony forming unit (CFU) per volume filtered.
1.2 This test method has been used successfully with temperate fresh and marine ambient waters, and wastewaters. It is the user’s
responsibility to ensure the validity of this test method for waters of other types.
1.3 The values stated in SI units are to be regarded as standard.
1.4 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, health, and environmental practices and determine the applicability of
regulatory limitations prior to use. For specific hazard statements, see Section 9.
1.5 This international standard was developed in accordance with internationally recognized principles on standardization
established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued
by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
2. Referenced Documents
2
2.1 ASTM Standards:
D1129 Terminology Relating to Water
D1193 Specification for Reagent Water
D3370 Practices for Sampling Water from Flowing Process Streams
3
D3870 Practice for Establishing Performance Characteristics for Colony Counting Methods in Microbiology (Withdrawn 2000)
D5465 Practices for Determining Microbial Colony Counts from Waters Analyzed by Plating Methods
3. Terminology
3.1 Definitions:
1
This test method is under the jurisdiction of ASTM Committee D19 on Water and is the direct responsibility of Subcommittee D19.24 on Water Microbiology.
Current edition approved April 1, 2019April 1, 2024. Published April 2019April 2024. Originally approved in 1993. Last previous edition approved in 20142019 as
D5392 – 14.D5392 – 19. DOI: 10.1520/D5392-19.10.1520/D5392-24.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM Standards
volume information, refer to the standard’s Document Summary page on the ASTM website.
3
The last approved version of this historical standard is referenced on www.astm.org.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 ----------------------
D5392 − 24
3.1.1 For definitions of terms used in this standard, refer to Terminology D1129.
3.2 Definitions of Terms Specific to This Standard:
3.2.1 Escherichia coli (E. coli), n—a species of bacteria that is a member of the total coliform group and known to originate in
the feces of warm-blooded animals.
3.3 Performance Characteristics (Practice D3870):
3.3.1 accuracy, n—the proportion of the observed count to the true density of a sample.
3.3.2 bias, n—the persistent positive or negative deviation of the average value of the test method from the assumed or accepted
true value.
3.3.3 precision, n—the degree of agreement of repeated measurements of the same parameter expressed quantitatively as the
standard deviation or as the 95 % confidence limits of the mean computed from the results of a series of controlled determinations.
3.3.4 specificity, n—the ability of a test method to select or distinguish, or both, the target bacteria in the same water sample; the
specificity characteristic of the method is usually reported as the percent of false positive and false negative results.
3.3.5 upper counting limit (UCL), n—that colony count above which there is an unacceptable counting error; the error may be due
to overcrowding or antibiosis.
4.
...
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