ASTM D6503-14
(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, 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 colony forming unit (CFU)/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 and health practices and determine the applicability of regulatory limitations prior to use.
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Designation: D6503 − 14
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 D2777Practice for Determination of Precision and Bias of
Applicable Test Methods of Committee D19 on Water
1.1 This test method covers a simple procedure for the
D3370Practices for Sampling Water from Flowing Process
detectionofenterococciinwaterandwastewater.Itisbasedon
2
Streams
IDEXX’spatentedDefinedSubstrateTechnology(DST). This
product, Enterolert, utilizes a nutrient indicator that fluoresces
3. Terminology
when metabolized. It can detect these bacteria at one colony
forming unit (CFU)/100 mL within 24 h. The presence of this
3.1 Definitions—For definitions of terms used in this test
microorganisminwaterisanindicationoffecalcontamination
method, refer to Terminology D1129.
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
responsibility of the user of this standard to establish appro-
3.2.3.1 Discussion—It is a qualitative value, “yes” or “no”
priate safety, health, and environmental practices and deter-
for reporting results.
mine the applicability of regulatory limitations prior to use.
3.2.4 Quanti-Tray, n—a system for the quantification of
1.5 This international standard was developed in accor-
enterococci.
dance with internationally recognized principles on standard-
3.2.4.1 Discussion—It consists of a sealer and trays which
ization established in the Decision on Principles for the
have multi-wells and can enumerate up to 2419 MPN/100 mL
Development of International Standards, Guides and Recom-
without dilution.
mendations issued by the World Trade Organization Technical
Barriers to Trade (TBT) Committee.
3.2.5 snap pack, n—apackagecontainingEnterolertreagent
for testing 100-mL sample either in the P/A format or
2. Referenced Documents
2
quantitatively, with the Quanti-Tray system.
3
2.1 ASTM Standards:
D1129Terminology Relating to Water
4. Summary of Test Method
D1193Specification for Reagent Water
4.1 Thistestmethodisusedforthedetectionofenterococci,
such as E. faecium, E. faecalis in drinking water, source water,
1
This test method is under the jurisdiction ofASTM Committee D19 on Water recreational waters (marine water and fresh), wastewaters, and
and is the direct responsibility of Subcommittee D19.24 on Water Microbiology.
bottled water. When the reagent is added to the sample and
Current edition approved Aug. 1, 2014. Published October 2014. Originally
incubatedat41 60.5°Cfor24handupto28h,Enterolertcan
approved in 1999. Last previous edition approved in 2009 as D6503–99 (2009).
detect these bacteria at 1 MPN/100 mL. Fluorescence is
DOI: 10.1520/D6503-14.
2
Trademark of IDEXX Laboratories, One Idexx Dr., Westbrook, ME 04092.
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
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D6503 − 14
5. Significance and Use 11.2 Qualitycontrolshouldbeconductedoneachnewlotof
Enterolert. See package insert for the recommended quality
5.1 This test provides an
...
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 − 99 (Reapproved 2009) D6503 − 14
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
2
IDEXX’sIDEXX’s 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)/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’suser’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 and health practices and determine the applicability of regulatory
limitations prior to use.
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 Closed Conduits
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 (366(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 isare present or absent in a water sample. It is a qualitative
value, “yes” or “no” for reporting results.
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 May 1, 2009Aug. 1, 2014. Published June 2009October 2014. Originally approved in 1999. Last previous edition approved in 20052009 as
D6503 – 99 (2005).(2009). DOI: 10.1520/D6503-99R09. 10.1520/D6503-14.
2
Trademark of IDEXX Laboratories, One Idexx Dr., Westbrook, ME 04092.
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.
3.2.3.1 Discussion—
It is a qualitative value, “yes” or “no” for reporting results.
2
3.2.4 quanti-trayQuanti-Tray , n—a system for the quantification of enterococci. It consists of a sealer and trays which have
multi-wells and can enumerate up to 2000 CFU/100 mL without dilution.
3.2.4.1 Discussion—
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 ----------------------
D6503 − 14
It consists of a sealer and trays which have multi-wells and can enumerate up to 2419 MPN/100 mL without dilution.
3.2.5 snap pack, n—a package containing Enterolert reagent for testing 100-mL sample either in the P/A format or
2
quantitatively, that is,with the Quanti-Tray system).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 waters (marine water and fresh), wastewaters, and bottled water. When the reagent is added to the sample and
incubated at 41 6 0.5°C for 24 h and up to 28 h, Enterolert can detect these bacteria at 1 CFU/100MPN/100 mL. Fluorescence
is produced when enterococci metabolizes the nutrient indicator. Enterolert
...
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