ASTM D6503-19
(Test Method)Standard Test Method for Enterococci in Water Using Enterolert
Standard Test Method for Enterococci in Water Using Enterolert
SIGNIFICANCE AND USE
5.1 This test provides an easy and reliable method for the detection of enterococci in water within 24 h. For recreational water (fresh and marine) testing is performed to insure areas are safe for swimming. Enterolert also can be used for testing bottled water, wastewater, ground water, and drinking water.
SCOPE
1.1 This test method covers a simple procedure for the detection of enterococci in water and wastewater. It is based on IDEXX’s patented Defined Substrate Technology (DST).2 This product, Enterolert, utilizes a nutrient indicator that fluoresces when metabolized. It can detect these bacteria at one most probable number (MPN)/100 mL within 24 h. The presence of this microorganism in water is an indication of fecal contamination and the possible presence of enteric pathogens.
1.2 This test method can be used successfully with drinking water, source water, recreational (fresh and marine) water, wastewater, and bottled water. It is the user’s responsibility to ensure the validity of this test method for waters of untested matrices.
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this 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.
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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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: D6503 − 19
Standard Test Method for
1,2
Enterococci in Water Using Enterolert
This standard is issued under the fixed designation D6503; 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
D2777Practice for Determination of Precision and Bias of
1.1 This test method covers a simple procedure for the
Applicable Test Methods of Committee D19 on Water
detectionofenterococciinwaterandwastewater.Itisbasedon
2 D3370Practices for Sampling Water from Flowing Process
IDEXX’spatentedDefinedSubstrateTechnology(DST). This
Streams
product, Enterolert, utilizes a nutrient indicator that fluoresces
when metabolized. It can detect these bacteria at one most
3. Terminology
probable number (MPN)/100 mLwithin 24 h. The presence of
3.1 Definitions—For definitions of terms used in this test
this microorganism in water is an indication of fecal contami-
method, refer to Terminology D1129.
nation and the possible presence of enteric pathogens.
3.2 Definitions of Terms Specific to This Standard:
1.2 This test method can be used successfully with drinking
3.2.1 enterococci, n—a gram positive bacteria possessing
water, source water, recreational (fresh and marine) water,
the enzyme β-D-glucosidase, which cleaves the nutrient indi-
wastewater, and bottled water. It is the user’s responsibility to
cator and produces fluorescence under a long wave length
ensure the validity of this test method for waters of untested
(365–366 nm) ultraviolet (UV) light.
matrices.
3.2.2 most probable number (MPN), n—a statistical method
1.3 The values stated in SI units are to be regarded as
for determining bacterial density based on the Poisson distri-
standard. No other units of measurement are included in this
bution.
standard.
3.2.3 presence-absence, n—a term used to indicate if en-
1.4 This standard does not purport to address all of the
terococci are present or absent in a water sample.
safety concerns, if any, associated with its use. It is the
3.2.3.1 Discussion—It is a qualitative value, “yes” or “no”
responsibility of the user of this standard to establish appro-
for reporting results.
priate safety, health, and environmental practices and deter-
3.2.4 Quanti-Tray, n—a system for the quantification of
mine the applicability of regulatory limitations prior to use.
2
enterococci.
1.5 This international standard was developed in accor-
3.2.4.1 Discussion—It consists of a sealer and trays which
dance with internationally recognized principles on standard-
have multi-wells and can enumerate up to 2419 MPN/100 mL
ization established in the Decision on Principles for the
without dilution using the Quanti-Tray 2000 tray or it can
Development of International Standards, Guides and Recom-
detect up to 200.5 MPN/100 mL using the 51-well Quanti-
mendations issued by the World Trade Organization Technical
Tray.
Barriers to Trade (TBT) Committee.
3.2.5 snap pack, n—apackagecontainingEnterolertreagent
2. Referenced Documents
for testing 100-mL sample either in the P/A format or
3
quantitatively, with the Quanti-Tray system.
2.1 ASTM Standards:
D1129Terminology Relating to Water
4. Summary of Test Method
4.1 Thistestmethodisusedforthedetectionofenterococci,
1 such as E. faecium, E. faecalis in drinking water, source water,
This test method is under the jurisdiction ofASTM Committee D19 on Water
and is the direct responsibility of Subcommittee D19.24 on Water Microbiology.
recreational waters (marine water and fresh), wastewaters, and
Current edition approved April 1, 2019. Published May 2019. Originally
bottled water. When the reagent is added to the sample and
approved in 1999. Last previous edition approved in 2014 as D6503–14. DOI:
incubatedat41 60.5°Cfor24handupto28h,Enterolertcan
10.1520/D6503-19.
2
detect these bacteria at 1 MPN/100 mL. Fluorescence is
Enterolert and Quanti-Tray are trademarks of IDEXX Laboratories, Inc., in
Westbrook, ME.
produced when enterococci metabolizes the nutrient indicator.
3
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
Enterolert can be used as a presence-absence test or for
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
quantification (5-tube, 10-tube MPN, 15-tube serial dilution or
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website. the Quanti-Tray system).
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700
...
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: D6503 − 14 D6503 − 19
Standard Test Method for
1,2
Enterococci in Water Using Enterolert
This standard is issued under the fixed designation D6503; 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 covers a simple procedure for the detection of enterococci in water and wastewater. It is based on IDEXX’s
2
patented Defined Substrate Technology (DST). This product, Enterolert, utilizes a nutrient indicator that fluoresces when
metabolized. It can detect these bacteria at one colony forming unit (CFU)/100most probable number (MPN)/100 mL within 24
h. The presence of this microorganism in water is an indication of fecal contamination and the possible presence of enteric
pathogens.
1.2 This test method can be used successfully with drinking water, source water, recreational (fresh and marine) water,
wastewater, and bottled water. It is the user’s responsibility to ensure the validity of this test method for waters of untested matrices.
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this 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.
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
3
2.1 ASTM Standards:
D1129 Terminology Relating to Water
D1193 Specification for Reagent Water
D2777 Practice for Determination of Precision and Bias of Applicable Test Methods of Committee D19 on Water
D3370 Practices for Sampling Water from Flowing Process Streams
3. Terminology
3.1 Definitions—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 enterococci, n—a gram positive bacteria possessing the enzyme β-D-glucosidase, which cleaves the nutrient indicator and
produces fluorescence under a long wave length (365–366 nm) ultraviolet (UV) light.
3.2.2 most probable number (MPN), n—a statistical method for determining bacterial density based on the Poisson distribution.
3.2.3 presence-absence, n—a term used to indicate if enterococci are present or absent in a water sample.
3.2.3.1 Discussion—
It is a qualitative value, “yes” or “no” for reporting results.
2
3.2.4 Quanti-Tray, n—a system for the quantification of enterococci.
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 Aug. 1, 2014April 1, 2019. Published October 2014May 2019. Originally approved in 1999. Last previous edition approved in 20092014 as
D6503 – 99 (2009).D6503 – 14. DOI: 10.1520/D6503-14.10.1520/D6503-19.
2
Trademark Enterolert and Quanti-Tray are trademarks of IDEXX Laboratories, One Idexx Dr., Westbrook, ME 04092. Inc., in Westbrook, ME.
3
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.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 ----------------------
D6503 − 19
3.2.4.1 Discussion—
It consists of a sealer and trays which have multi-wells and can enumerate up to 2419 MPN/100 mL without dilution.dilution using
the Quanti-Tray 2000 tray or it can detect up to 200.5 MPN/100 mL using the 51-well Quanti-Tray.
3.2.5 snap pack, n—a package containing Enterolert reagent for testing 100-mL sample either in the P/A format or
quantitatively, with the Quanti-Tray system.
4. Summary of Test Method
4.1 This test method is used for the detection of enterococci, such as E. faecium, E. faecalis in drinking water, source water,
recreational
...
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: D6503 − 19
Standard Test Method for
1,2
Enterococci in Water Using Enterolert
This standard is issued under the fixed designation D6503; 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
D2777 Practice for Determination of Precision and Bias of
1.1 This test method covers a simple procedure for the
Applicable Test Methods of Committee D19 on Water
detection of enterococci in water and wastewater. It is based on
2 D3370 Practices for Sampling Water from Flowing Process
IDEXX’s patented Defined Substrate Technology (DST). This
Streams
product, Enterolert, utilizes a nutrient indicator that fluoresces
when metabolized. It can detect these bacteria at one most
3. Terminology
probable number (MPN)/100 mL within 24 h. The presence of
3.1 Definitions—For definitions of terms used in this test
this microorganism in water is an indication of fecal contami-
method, refer to Terminology D1129.
nation and the possible presence of enteric pathogens.
3.2 Definitions of Terms Specific to This Standard:
1.2 This test method can be used successfully with drinking
3.2.1 enterococci, n—a gram positive bacteria possessing
water, source water, recreational (fresh and marine) water,
the enzyme β-D-glucosidase, which cleaves the nutrient indi-
wastewater, and bottled water. It is the user’s responsibility to
cator and produces fluorescence under a long wave length
ensure the validity of this test method for waters of untested
(365–366 nm) ultraviolet (UV) light.
matrices.
3.2.2 most probable number (MPN), n—a statistical method
1.3 The values stated in SI units are to be regarded as
for determining bacterial density based on the Poisson distri-
standard. No other units of measurement are included in this
bution.
standard.
3.2.3 presence-absence, n—a term used to indicate if en-
1.4 This standard does not purport to address all of the
terococci are present or absent in a water sample.
safety concerns, if any, associated with its use. It is the
3.2.3.1 Discussion—It is a qualitative value, “yes” or “no”
responsibility of the user of this standard to establish appro-
for reporting results.
priate safety, health, and environmental practices and deter-
3.2.4 Quanti-Tray, n—a system for the quantification of
mine the applicability of regulatory limitations prior to use.
2
enterococci.
1.5 This international standard was developed in accor-
3.2.4.1 Discussion—It consists of a sealer and trays which
dance with internationally recognized principles on standard-
have multi-wells and can enumerate up to 2419 MPN/100 mL
ization established in the Decision on Principles for the
without dilution using the Quanti-Tray 2000 tray or it can
Development of International Standards, Guides and Recom-
detect up to 200.5 MPN/100 mL using the 51-well Quanti-
mendations issued by the World Trade Organization Technical
Tray.
Barriers to Trade (TBT) Committee.
3.2.5 snap pack, n—a package containing Enterolert reagent
2. Referenced Documents
for testing 100-mL sample either in the P/A format or
3
quantitatively, with the Quanti-Tray system.
2.1 ASTM Standards:
D1129 Terminology Relating to Water
4. Summary of Test Method
4.1 This test method is used for the detection of enterococci,
1
such as E. faecium, E. faecalis in drinking water, source water,
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. recreational waters (marine water and fresh), wastewaters, and
Current edition approved April 1, 2019. Published May 2019. Originally
bottled water. When the reagent is added to the sample and
approved in 1999. Last previous edition approved in 2014 as D6503 – 14. DOI:
incubated at 41 6 0.5°C for 24 h and up to 28 h, Enterolert can
10.1520/D6503-19.
2
detect these bacteria at 1 MPN/100 mL. Fluorescence is
Enterolert and Quanti-Tray are trademarks of IDEXX Laboratories, Inc., in
Westbrook, ME.
produced when enterococci metabolizes the nutrient indicator.
3
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
Enterolert can be used as a presence-absence test or for
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
quantification (5-tube, 10-tube MPN, 15-tube serial dilution or
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website. the Quanti-Tray system).
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 --------
...
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