ASTM D4412-84(2009)
(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
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.
It has been reported that Desulfovibrio 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 D 2777 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 and health practices and determine the applicability of regulatory limitations prior to use.
General Information
Relations
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: D4412 − 84 (Reapproved 2009)
StandardTest Methods for
Sulfate-Reducing Bacteria in Water and Water-Formed
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 2. Referenced Documents
1.1 Thesetestmethodscovertheprocedureforthedetection 2.1 ASTM Standards:
and enumeration by the most probable number (MPN) tech- D1129Terminology Relating to Water
nique of sulfate-reducing bacteria in water or water-formed D1193Specification for Reagent Water
deposits. D2777Practice for Determination of Precision and Bias of
Applicable Test Methods of Committee D19 on Water
1.2 Two media preparations are provided. MediumAwhich
2.2 APHA Standard:
is prepared with reagent grade water, and Medium B which is
Standard Methods for the Examination of Water and
prepared using the water to be sampled as the water source.
Wastewater, Fifteenth Edition
Medium B is offered for those special conditions where
sulfate-reducing bacterial strains have adapted to atypical
3. Terminology
non-fresh water environment.
3.1 Definitions—For definitions of terms used in these test
1.3 For the isolation and enumeration of thermophilic
methods, refer to Terminology D1129.
sulfate-reducingbacteriaencounteredinwatersassociatedwith
3.2 Definitions—For a description of the term MPN used in
oil and gas production, all broths, dilution blanks, and incuba-
these test methods, refer to literature.
tions must be maintained at temperatures of at least 45°C and
preferably within 5°C at the sample temperature.
4. Summary of Test Methods
1.4 Thesensitivityofthesetestmethodscanbeincreasedby
4.1 Water and water deposit samples and dilutions of these
purging the dilution blanks and tubes of media with nitrogen
samplesaredispensedintotubesofStarkey’smedium(AorB)
immediately prior to use.
followingfivetubeMPNprocedures.Thetubesaresealedwith
liquid paraffin, and incubated at 20°C for 21 days. Positive
1.5 The analyst should be aware that adequate collaborative
reactions are indicated by the deposit of a black precipitate.
data for precision and bias statements as required by Practice
D2777 are not provided. See Section 11 for details.
5. Significance and Use
1.6 The values stated in SI units are to be regarded as
5.1 Sulfate-reducing bacteria are widely distributed in ma-
standard. No other units of measurement are included in this
rine and fresh water muds which, in consequence, frequently
standard.
are laden with the hydrogen sulfide produced by these organ-
1.7 This standard does not purport to address all of the
isms during dissimilatory sulfate reduction.
safety concerns, if any, associated with its use. It is the
5.2 It has been reported that Desulfovibrio can form as
responsibility of the user of this standard to establish appro-
much as 10 g of sulfide per litre during active multiplication.
priate safety and health practices and determine the applica-
Sulfate-reducing bacteria can cause the external or internal
bility of regulatory limitations prior to use.
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
These test methods are under the jurisdiction of ASTM Committee D19 on Standards volume information, refer to the standard’s Document Summary page on
Water and are the direct responsibility of Subcommittee D19.24 on Water Micro- the ASTM website.
biology. Available from American Public Health Association, 1015 18th St. N.W.,
Current edition approved May 1, 2009. Published June 2009. Originally Washington, DC 20036.
approved in 1984. Last previous edition approved in 2002 as D4412–84 (2002). Bonde,G.J.,“BacterialIndicatorsofWaterPollution,” A Study of Quantitative
DOI: 10.1520/D4412-84R09.
Estimation, Teknisk Forlag, Copenhagen, 1963.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
D4412 − 84 (Reapproved 2009)
corrosion of water or wastewater pipelines and pipelines for 7.4.1 Watercollectedfromthesamplecollectionsiteisused
petroleum and natural gas. The formation of galvanic cells by to prepare the medium outlined in 7.3. The water sample is
massive growth of sulfate-reducing bacteria under suitable filteredtoremoveparticulates(1.2µmmembranefilter)andthe
conditionsmakesthecorrosionmuchworsethanjusttheeffect pH is recorded.
of the hydrogen sulfide on the metal or concrete. 7.4.1.1 After preparing the Medium B following 7.3.1,
7.3.2, and 7.3.3, and prior to dispensing, check and adjust pH,
6. Apparatus and Materials
if necessary to that of the original water used, then filter
6.1 Anaerobic Incubator,20°C,ifavailable,orconventional sterilize the medium by passage through 0.2-µm filter and
asceptically dispense into presterilized tubes.
20°C incubator.
7.5 Hydrogen Sulfide Test Reagent :
NOTE 1—For thermophilic organisms use a 45°C incubator.
7.5.1 Ferric Chloride Stock Solution(FeCl ·6H O)— Dis-
3 2
6.2 Pipets, sterile, 1 mLand 10 mL, “calibrated” to deliver.
solve 13.5 g of ferric chloride in a mixture of 250 mLof water
6.3 Test Tubes, with close fitting or airtight caps; 16 by 150
and 250 mL of HCl (sp gr 1.19). Store in an airtight amber
mm and 20 by 150 mm.
container. Prepare fresh monthly.
6.4 Test Tube Racks, of sufficient size to contain 16 and 7.5.2 p-Aminodimethylaniline Dihydrochloride Stock Solu-
20-mm tubes. tion
p-Aminodimethylaniline dihydrochloride 1.0 g
7. Reagents
(C H N ·2HCl)
8 12 2
HCl (6 N) 500 mL
7.1 Purity of Reagents—Reagent grade chemicals shall be
Dissolve1gof p-aminodimethylaniline dihydrochloride in
used in all tests. Unless otherwise indicated, it is intended that
500 mL of 6 N HCl. Store for up to 1 month in an amber
all reagents conform to the specifications of the Committee on
airtight container.
Analytical Reagents of theAmerican Chemical Society, when
such specifications are available.
7.6 Liquid Paraffın— Heavy, sterile, or sterile mineral oil.
7.2 Purity of Water—Unlessotherwiseindicated,references
7.7 Buffered Dilution Water—Stock Solution
to water shall be understood to mean Reagent Water Type II
7.7.1 Dissolve34.0gofKH PO in500mLofwater,adjust
2 4
conforming to Specification D1193. In addition, reagent water
pHto7.2with1NNaOHanddiluteto1Lwith
...
Questions, Comments and Discussion
Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.