ASTM D8429-21

Standard Test Method for <emph type="bdit">Legionella pneumophila</emph> in Water Samples Using Legiolert

SIGNIFICANCE AND USE
5.1 This test provides an easy and reliable method for the detection of L. pneumophila in potable and non-potable waters in 7 days.  
5.2 Routine monitoring for L. pneumophila determines whether implemented control measures are effective, such as those outlined in a water safety program (2).  
5.2.1 Water system management is necessary to maintain L. pneumophila concentrations below hazardous levels. Through routine measurement of L. pneumophila levels, a monitoring program can ensure that control measures are effective and implemented when necessary in response to increasing levels. Water samples may be examined for L. pneumophila during epidemiological investigations as part of local authority, industrial, or hospital programs, or in order to validate treatment control methods. Routine sampling could also be carried out based on risk assessments or on local, state, or federal requirements or guidelines.
SCOPE
1.1 This test method describes a simple procedure for the detection of Legionella pneumophila (L. pneumophila) in potable water and non-potable waters (cooling towers, for example). This procedure describes a liquid culture method based on a bacterial enzyme technology. The detection of L. pneumophila is signaled through the utilization of a substrate present in the Legiolert reagent. L. pneumophila cells grow rapidly and reproduce using the rich supply of amino acids, vitamins and other nutrients present in the Legiolert reagent. Actively growing strains of L. pneumophila use the added substrate to produce a brown color indicator or produce turbid growth with or without brown coloration. Legiolert can detect this bacterial species at the following minimum concentrations based on the protocol employed:  
1.1.1 Potable Water:  
1.1.1.1 ≥1 organism / 100 mL at 7 days for 100 mL potable protocol.
1.1.1.2 ≥1 organism / 10 mL at 7 days for 10 mL potable protocol.  
1.1.2 Non-potable Water:  
1.1.2.1 ≥1 organism / 1.0 mL at 7 days for 1.0 mL non-potable protocol.
1.1.2.2 ≥1 organism / 0.1 mL at 7 days for 0.1 mL non-potable protocol.  
1.1.3 This test method can be used for potable (drinking) waters and non-potable waters such as cooling tower waters (1).3 It is the user’s responsibility to ensure the validity of this test method for waters of untested matrices.  
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, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.4 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.

General Information

• Status: Published
• Publication Date: 30-Nov-2021
• ICS: 07.100.20 - Microbiology of water
• ICS: 13.060.45 - Examination of water in general
• Technical Committee: D19 - Water
• Drafting Committee: D19.24 - Water Microbiology

Standards Content (Sample)

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: D8429 − 21
Standard Test Method for
1,2
Legionella pneumophila in Water Samples Using Legiolert
This standard is issued under the fixed designation D8429; 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 priate safety, health, and environmental practices and deter-
mine the applicability of regulatory limitations prior to use.
1.1 This test method describes a simple procedure for the
1.4 This international standard was developed in accor-
detection of Legionella pneumophila (L. pneumophila)in
dance with internationally recognized principles on standard-
potable water and non-potable waters (cooling towers, for
ization established in the Decision on Principles for the
example). This procedure describes a liquid culture method
Development of International Standards, Guides and Recom-
based on a bacterial enzyme technology. The detection of L.
mendations issued by the World Trade Organization Technical
pneumophila is signaled through the utilization of a substrate
Barriers to Trade (TBT) Committee.
present in the Legiolert reagent. L. pneumophila cells grow
rapidly and reproduce using the rich supply of amino acids,
2. Referenced Documents
vitamins and other nutrients present in the Legiolert reagent.
4
2.1 ASTM Standards:
Actively growing strains of L. pneumophila use the added
D1129 Terminology Relating to Water
substrate to produce a brown color indicator or produce turbid
D1193 Specification for Reagent Water
growth with or without brown coloration. Legiolert can detect
D2777 Practice for Determination of Precision and Bias of
this bacterial species at the following minimum concentrations
Applicable Test Methods of Committee D19 on Water
based on the protocol employed:
D3370 Practices for Sampling Water from Flowing Process
1.1.1 Potable Water:
Streams
1.1.1.1 ≥1 organism / 100 mL at 7 days for 100 mL potable
D5952 Guide for the Inspection of Water Systems for
protocol.
Legionella and the Investigation of Possible Outbreaks of
1.1.1.2 ≥1 organism / 10 mL at 7 days for 10 mL potable
Legionellosis (Legionnaires’ Disease or Pontiac Fever)
protocol.
D6503 Test Method for Enterococci in Water Using Enter-
1.1.2 Non-potable Water:
olert
1.1.2.1 ≥1 organism / 1.0 mL at 7 days for 1.0 mL
non-potable protocol.
3. Terminology
1.1.2.2 ≥1 organism / 0.1 mL at 7 days for 0.1 mL
3.1 Definitions:
non-potable protocol.
3.1.1 For definitions of terms used in this standard, refer to
1.1.3 This test method can be used for potable (drinking)
Terminology D1129.
waters and non-potable waters such as cooling tower waters
3.1.2 Legionella, n—a bacterial genus containing over 50
3
(1). It is the user’s responsibility to ensure the validity of this
species and at least 71 serogroups; abbreviated to the first
test method for waters of untested matrices.
initialwhenusedrelatedlywithaspeciesname,forexample,L.
1.2 The values stated in SI units are to be regarded as
pneumophila.
standard. No other units of measurement are included in this
3.1.3 most probable number (MPN), n—a statistical method
standard.
for determining bacterial concentration based on the Poisson
1.3 This standard does not purport to address all of the
distribution.
safety concerns, if any, associated with its use. It is the
3.2 Definitions of Terms Specific to This Standard:
responsibility of the user of this standard to establish appro-
3.2.1 Legionella pneumophila, n—in the context of this
method,apathogenicgram-negativebacteriathatproducesany
degree of brown coloration or turbid growth with or without
1
Legiolert is a trademark of IDEXX Laboratories, Inc., Westbrook, ME.
brown coloration in the presence of the Legiolert reagent.
2
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.
4
Current edition approved Dec. 1, 2021. Published December 2021. DOI: For referenced ASTM standards, visit the ASTM website, www.astm.org, or
10.1520/D8429-21. contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM
3
The boldface numbers in parentheses refer to a list of references at the end of Standards volume information, refer to the standard’s Document Summary page on
this standard. the ASTM website.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
D8429 − 21
3
...