ASTM D5392-14

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 and health practices and determine the applicability of regulatory limitations prior to use. For specific hazard statements, see Section 9.

General Information

• Status: Historical
• Publication Date: 14-Aug-2014
• ICS: 07.100.20 - Microbiology of water
• Technical Committee: D19 - Water
• Drafting Committee: D19.24 - Water Microbiology

Relations

• Replaces: ASTM D5392-93(2006) - Standard Test Method for Isolation and Enumeration ofEscherichia Coli in Water by the Two-Step Membrane Filter Procedure
• Effective Date: 15-Aug-2014
• Replaced By: ASTM D5392-19 - Standard Test Method for Isolation and Enumeration of <emph type="bdit">Escherichia coli</emph> in Water by the Two-Step Membrane Filter Procedure
• Effective Date: 01-Apr-2019
• Referred By: ASTM D8506-23 - Standard Guide for Microbial Contamination and Biodeterioration in Turbine Oils and Turbine Oil Systems
• Effective Date: 15-Aug-2014
• Referred By: ASTM D5465-16(2020) - Standard Practices for Determining Microbial Colony Counts from Waters Analyzed by Plating Methods
• Effective Date: 15-Aug-2014

Standards Content (Sample)

NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation: D5392 − 14
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.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope D1193Specification for Reagent Water
D3370Practices for Sampling Water from Closed Conduits
1.1 This test method describes a membrane filter (MF)
D3870PracticeforEstablishingPerformanceCharacteristics
procedure for the detection and enumeration of Escherichia
for Colony Counting Methods in Microbiology (With-
coli, a bacterium found exclusively in the feces of humans and
3
drawn 2000)
other warm-blooded animals. The presence of these microor-
D5465Practices for Determining Microbial Colony Counts
ganisms in water is an indication of fecal pollution and the
from Waters Analyzed by Plating Methods
possible presence of enteric pathogens. These bacteria are
foundinwaterandwastewaterinawiderangeofdensities.The
3. Terminology
detection limit of this procedure is one colony forming unit
(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 test method, refer
temperatefreshandmarineambientwaters,andwastewaters.It
to Terminology D1129.
is the user’s responsibility to ensure the validity of this test
3.2 Definitions of Terms Specific to This Standard:
method for waters of other types.
3.2.1 Escherichia coli (E. coli), n—aspeciesofbacteriathat
1.3 The values stated in SI units are to be regarded as isamemberofthetotalcoliformgroupandknowntooriginate
standard.
in the feces of warm-blooded animals.
1.4 This standard does not purport to address all of the
3.3 Performance Characteristics (Practice D3870):
safety concerns, if any, associated with its use. It is the
3.3.1 accuracy, n—the proportion of the observed count to
responsibility of the user of this standard to establish appro-
the true density of a sample.
priate safety, health, and environmental practices and deter-
3.3.2 bias, n—the persistent positive or negative deviation
mine the applicability of regulatory limitations prior to use.
of the average value of the test method from the assumed or
For specific hazard statements, see Section 9.
accepted true value.
1.5 This international standard was developed in accor-
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
D1129Terminology Relating to Water
reported as the percent of false positive and false negative
results.
1
This test method is under the jurisdiction ofASTM 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 Aug. 15, 2014. Published August 2014. Originally
may be due to overcrowding or antibiosis.
approved in 1993. Last previous edition approved in 2006 as D5392–93 (2006).
DOI: 10.1520/D5392-14.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at [e-mail]. 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 − 14
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 provides a direct count of
bacterial colonies developing on the surface of the filter when 7.3 Counting Device, hand tally or electronic.
placed on a selective nutrient medium. The water sample is
7.4 Pipe
...

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 − 93 (Reapproved 2006) D5392 − 14
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 and health practices and determine the applicability of regulatory
limitations prior to use. For specific hazard statements, see Section 9.
2. Referenced Documents
2
2.1 ASTM Standards:
D1129 Terminology Relating to Water
D1193 Specification for Reagent Water
D3370 Practices for Sampling Water from Closed Conduits
3
D3870 Practice for Establishing Performance Characteristics for Colony Counting Methods in Microbiology (Withdrawn 2000)
D5465 Practice for Determining Microbial Colony Counts from Waters Analyzed by Plating Methods
3. Terminology
3.1 Definitions—For definitions of terms used in this test method, refer to Terminology D1129.
3.1 Definitions:
3.1.1 For definitions of terms used in this test method, refer to Terminology D1129.
3.2 Definitions of Terms Specific to This Standard:
3.2.1 Escherichia coli (E. coli)—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—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.
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 July 1, 2006Aug. 15, 2014. Published July 2006August 2014. Originally approved in 1993. Last previous edition approved in 20002006 as
D5392 – 93 (2000).(2006). DOI: 10.1520/D5392-93R06. 10.1520/D5392-14.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at [e-mail]. 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 − 14
3.3.2 bias—the persistent positive or negative deviation of the average value of the test method from the assumed or accepted
true value.
3.3.4 specificity—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)—(UCL), n—that colony count above which there is an unacceptable counting error; the error
may be due to overcrowding or antibiosis.
3.3.5 accuracy—the proportion of the observed count to th
...