ASTM D4412-19
(Test Method)Standard Test Methods for Sulfate-Reducing Bacteria in Water and Water-Formed Deposits
Standard Test Methods for Sulfate-Reducing Bacteria in Water and Water-Formed Deposits
SIGNIFICANCE AND USE
5.1 Sulfate-reducing bacteria are widely distributed in marine and fresh water muds which, in consequence, frequently are laden with the hydrogen sulfide produced by these organisms during dissimilatory sulfate reduction.
5.2 It has been reported that Desulfovibrio spp. can form as much as 10 g of sulfide per litre during active multiplication. Sulfate-reducing bacteria can cause the external or internal corrosion of water or wastewater pipelines and pipelines for petroleum and natural gas. The formation of galvanic cells by massive growth of sulfate-reducing bacteria under suitable conditions makes the corrosion much worse than just the effect of the hydrogen sulfide on the metal or concrete.
SCOPE
1.1 These test methods cover the procedure for the detection and enumeration by the most probable number (MPN) technique of sulfate-reducing bacteria in water or water-formed deposits.
1.2 Two media preparations are provided. Medium A which is prepared with reagent grade water, and Medium B which is prepared using the water to be sampled as the water source. Medium B is offered for those special conditions where sulfate-reducing bacterial strains have adapted to atypical non-fresh water environment.
1.3 For the isolation and enumeration of thermophilic sulfate-reducing bacteria encountered in waters associated with oil and gas production, all broths, dilution blanks, and incubations must be maintained at temperatures of at least 45°C and preferably within 5°C at the sample temperature.
1.4 The sensitivity of these test methods can be increased by purging the dilution blanks and tubes of media with nitrogen immediately prior to use.
1.5 The analyst should be aware that adequate collaborative data for precision and bias statements as required by Practice D2777 are not provided. See Section 11 for details.
1.6 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.7 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.
1.8 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: D4412 − 19
Standard Test Methods for
Sulfate-Reducing Bacteria in Water and Water-Formed
1
Deposits
This standard is issued under the fixed designation D4412; 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* mendations issued by the World Trade Organization Technical
Barriers to Trade (TBT) Committee.
1.1 Thesetestmethodscovertheprocedureforthedetection
and enumeration by the most probable number (MPN) tech-
2. Referenced Documents
nique of sulfate-reducing bacteria in water or water-formed
2
2.1 ASTM Standards:
deposits.
D1129Terminology Relating to Water
1.2 Two media preparations are provided. MediumAwhich
D1193Specification for Reagent Water
is prepared with reagent grade water, and Medium B which is
D2777Practice for Determination of Precision and Bias of
prepared using the water to be sampled as the water source.
Applicable Test Methods of Committee D19 on Water
Medium B is offered for those special conditions where
D6503Test Method for Enterococci in Water Using Enter-
sulfate-reducing bacterial strains have adapted to atypical
olert
non-fresh water environment.
2.2 Other Standards:
1.3 For the isolation and enumeration of thermophilic
Standard Methods 9221Multiple-Tube Fermentation Tech-
3
sulfate-reducingbacteriaencounteredinwatersassociatedwith
nique for Members of the Coliform Group
oil and gas production, all broths, dilution blanks, and incuba-
tions must be maintained at temperatures of at least 45°C and
3. Terminology
preferably within 5°C at the sample temperature.
3.1 Definitions—For definitions of terms used in this
1.4 Thesensitivityofthesetestmethodscanbeincreasedby
standard, refer to Terminology D1129.
purging the dilution blanks and tubes of media with nitrogen
3.1.1 most probable number, n—statistical method for de-
immediately prior to use.
termining bacterial density based on the Poisson distribution.
D6503
1.5 The analyst should be aware that adequate collaborative
data for precision and bias statements as required by Practice
3.2 Acronyms:
D2777 are not provided. See Section 11 for details.
3.2.1 SRB, n—sulfate-reducing bacteria
1.6 The values stated in SI units are to be regarded as
4. Summary of Test Methods
standard. No other units of measurement are included in this
standard.
4.1 Water and water deposit samples and dilutions of these
samplesaredispensedintotubesofStarkey’smedium(AorB)
1.7 This standard does not purport to address all of the
followingfivetubeMPNprocedures.Thetubesaresealedwith
safety concerns, if any, associated with its use. It is the
4
liquid paraffin, and incubated at 20°C for 21 days. Positive
responsibility of the user of this standard to establish appro-
reactions are indicated by the deposit of a black precipitate.
priate safety, health, and environmental practices and deter-
mine the applicability of regulatory limitations prior to use.
5. Significance and Use
1.8 This international standard was developed in accor-
dance with internationally recognized principles on standard- 5.1 Sulfate-reducing bacteria are widely distributed in ma-
ization established in the Decision on Principles for the rine and fresh water muds which, in consequence, frequently
Development of International Standards, Guides and Recom-
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
1
These test methods are under the jurisdiction of ASTM Committee D19 on contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Water and are the direct responsibility of Subcommittee D19.24 on Water Micro- Standards volume information, refer to the standard’s Document Summary page on
biology. the ASTM website.
3
Current edition approved Dec. 1, 2019. Published December 2019. Originally Available from Standard Methods, http://standardmethods.org.
4
approved in 1984. Last previous edition approved in 2015 as D4412–15. DOI: Starkey, R. L., “Characteristics and Cultivation of Sulfate-Reducing Bacteria,”
10.1520/D4412-19. Journal of the American Water Works Association, Vol 40, 1948, pp. 1291–1298.
*A Summary of Changes section appears at the end of this standard
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 ----------------------
D4412 − 19
are laden with the hydrogen sulfide produced by these organ- 7.3.4 pH of medium should be 7.2 after autoclave
isms during dissimilatory sulfat
...
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: D4412 − 15 D4412 − 19
Standard Test Methods for
Sulfate-Reducing Bacteria in Water and Water-Formed
1
Deposits
This standard is issued under the fixed designation D4412; 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 These test methods cover the procedure for the detection and enumeration by the most probable number (MPN) technique
of sulfate-reducing bacteria in water or water-formed deposits.
1.2 Two media preparations are provided. Medium A which is prepared with reagent grade water, and Medium B which is
prepared using the water to be sampled as the water source. Medium B is offered for those special conditions where
sulfate-reducing bacterial strains have adapted to atypical non-fresh water environment.
1.3 For the isolation and enumeration of thermophilic sulfate-reducing bacteria encountered in waters associated with oil and
gas production, all broths, dilution blanks, and incubations must be maintained at temperatures of at least 45°C and preferably
within 5°C at the sample temperature.
1.4 The sensitivity of these test methods can be increased by purging the dilution blanks and tubes of media with nitrogen
immediately prior to use.
1.5 The analyst should be aware that adequate collaborative data for precision and bias statements as required by Practice D2777
are not provided. See Section 11 for details.
1.6 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.7 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 safety, health, and healthenvironmental practices and determine the
applicability of regulatory limitations prior to use.
1.8 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
D2777 Practice for Determination of Precision and Bias of Applicable Test Methods of Committee D19 on Water
D6503 Test Method for Enterococci in Water Using Enterolert
2.2 Other Standards:
3
Standard Methods 9221 Multiple-Tube Fermentation Technique for Members of the Coliform Group
3. Terminology
3.1 Definitions—For definitions of terms used in this standard, refer to Terminology D1129.
3.1.1 most probable number, n—statistical method for determining bacterial density based on the Poisson distribution. D6503
3.2 Acronyms:
1
These test methods are under the jurisdiction of ASTM Committee D19 on Water and are the direct responsibility of Subcommittee D19.24 on Water Microbiology.
Current edition approved July 15, 2015Dec. 1, 2019. Published August 2015December 2019. Originally approved in 1984. Last previous edition approved in 20092015
as D4412 – 84 (2009).D4412 – 15. DOI: 10.1520/D4412-15.10.1520/D4412-19.
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’sstandard’s Document Summary page on the ASTM website.
3
Available from Standard Methods, http://standardmethods.org.
*A Summary of Changes section appears at the end of this standard
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 ----------------------
D4412 − 19
3.2.1 SRB, n—sulfate-reducing bacteria
4. Summary of Test Methods
4.1 Water and water deposit samples and dilutions of these samples are dispensed into tubes of Starkey’sStarkey’s medium (A
4
or B) following five tube MPN procedures. The tubes are sealed with liquid paraffin, and incubated at 20°C for 21 days. Positive
reactions are indicated by the deposit of a black precipitate.
5. Significance and Use
5.1 Sulfate-reducing bacteria are widely distributed in marine and fresh water muds which, in consequence, frequently are laden
with the hydrogen sulfide produced
...
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: D4412 − 19
Standard Test Methods for
Sulfate-Reducing Bacteria in Water and Water-Formed
1
Deposits
This standard is issued under the fixed designation D4412; 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* mendations issued by the World Trade Organization Technical
Barriers to Trade (TBT) Committee.
1.1 These test methods cover the procedure for the detection
and enumeration by the most probable number (MPN) tech-
2. Referenced Documents
nique of sulfate-reducing bacteria in water or water-formed
2
2.1 ASTM Standards:
deposits.
D1129 Terminology Relating to Water
1.2 Two media preparations are provided. Medium A which
D1193 Specification for Reagent Water
is prepared with reagent grade water, and Medium B which is
D2777 Practice for Determination of Precision and Bias of
prepared using the water to be sampled as the water source.
Applicable Test Methods of Committee D19 on Water
Medium B is offered for those special conditions where
D6503 Test Method for Enterococci in Water Using Enter-
sulfate-reducing bacterial strains have adapted to atypical
olert
non-fresh water environment.
2.2 Other Standards:
1.3 For the isolation and enumeration of thermophilic
Standard Methods 9221 Multiple-Tube Fermentation Tech-
sulfate-reducing bacteria encountered in waters associated with 3
nique for Members of the Coliform Group
oil and gas production, all broths, dilution blanks, and incuba-
tions must be maintained at temperatures of at least 45°C and
3. Terminology
preferably within 5°C at the sample temperature.
3.1 Definitions—For definitions of terms used in this
1.4 The sensitivity of these test methods can be increased by
standard, refer to Terminology D1129.
purging the dilution blanks and tubes of media with nitrogen
3.1.1 most probable number, n—statistical method for de-
immediately prior to use.
termining bacterial density based on the Poisson distribution.
D6503
1.5 The analyst should be aware that adequate collaborative
data for precision and bias statements as required by Practice
3.2 Acronyms:
D2777 are not provided. See Section 11 for details.
3.2.1 SRB, n—sulfate-reducing bacteria
1.6 The values stated in SI units are to be regarded as
4. Summary of Test Methods
standard. No other units of measurement are included in this
standard.
4.1 Water and water deposit samples and dilutions of these
samples are dispensed into tubes of Starkey’s medium (A or B)
1.7 This standard does not purport to address all of the
following five tube MPN procedures. The tubes are sealed with
safety concerns, if any, associated with its use. It is the
4
liquid paraffin, and incubated at 20°C for 21 days. Positive
responsibility of the user of this standard to establish appro-
reactions are indicated by the deposit of a black precipitate.
priate safety, health, and environmental practices and deter-
mine the applicability of regulatory limitations prior to use.
5. Significance and Use
1.8 This international standard was developed in accor-
dance with internationally recognized principles on standard- 5.1 Sulfate-reducing bacteria are widely distributed in ma-
ization established in the Decision on Principles for the rine and fresh water muds which, in consequence, frequently
Development of International Standards, Guides and Recom-
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
1
These test methods are under the jurisdiction of ASTM Committee D19 on contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Water and are the direct responsibility of Subcommittee D19.24 on Water Micro- Standards volume information, refer to the standard’s Document Summary page on
biology. the ASTM website.
3
Current edition approved Dec. 1, 2019. Published December 2019. Originally Available from Standard Methods, http://standardmethods.org.
4
approved in 1984. Last previous edition approved in 2015 as D4412 – 15. DOI: Starkey, R. L., “Characteristics and Cultivation of Sulfate-Reducing Bacteria,”
10.1520/D4412-19. Journal of the American Water Works Association, Vol 40, 1948, pp. 1291–1298.
*A Summary of Changes section appears at the end of this standard
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 ----------------------
D4412 − 19
are laden with the hydrogen sulfide produced by these organ- 7.3.4 pH of medium should be 7.2 after autoclave
isms during dissimilatory sulfate reduction. sterilization, at 121°C for 15 min.
5.2 It has been reported that Desulfovibrio spp
...
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