ASTM_D_4412_-_15.pdf

Designation:D441215Standard Test Methods forSulfate-Reducing Bacteria in Water and Water-FormedDeposits1This standard is issued under the fixed designation D4412;the number immediately following the designation indicates the year oforiginal adoption or,in the case of revision,the year of last revision.A number in parentheses indicates the year of last reapproval.Asuperscript epsilon()indicates an editorial change since the last revision or reapproval.1.Scope*1.1 These test methods cover the procedure for the detectionand enumeration by the most probable number(MPN)tech-nique of sulfate-reducing bacteria in water or water-formeddeposits.1.2 Two media preparations are provided.Medium A whichis prepared with reagent grade water,and Medium B which isprepared using the water to be sampled as the water source.Medium B is offered for those special conditions wheresulfate-reducing bacterial strains have adapted to atypicalnon-fresh water environment.1.3 For the isolation and enumeration of thermophilicsulfate-reducing bacteria encountered in waters associated withoil and gas production,all broths,dilution blanks,and incuba-tions must be maintained at temperatures of at least 45C andpreferably within 5C at the sample temperature.1.4 The sensitivity of these test methods can be increased bypurging the dilution blanks and tubes of media with nitrogenimmediately prior to use.1.5 The analyst should be aware that adequate collaborativedata for precision and bias statements as required by PracticeD2777 are not provided.See Section 11 for details.1.6 The values stated in SI units are to be regarded asstandard.No other units of measurement are included in thisstandard.1.7 This standard does not purport to address all of thesafety concerns,if any,associated with its use.It is theresponsibility of the user of this standard to establish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use.2.Referenced Documents2.1 ASTM Standards:2D1129 Terminology Relating to WaterD1193 Specification for Reagent WaterD2777 Practice for Determination of Precision and Bias ofApplicable Test Methods of Committee D19 on WaterD6503 Test Method for Enterococci in Water Using Enter-olert2.2 Other Standards:Standard Methods 9221 Multiple-Tube Fermentation Tech-nique for Members of the Coliform Group33.Terminology3.1 DefinitionsFor definitions of terms used in thisstandard,refer to Terminology D1129.3.1.1 most probable number,nstatistical method for de-termining bacterial density based on the Poisson distribution.D65033.2 Acronyms:3.2.1 SRB,nsulfate-reducing bacteria4.Summary of Test Methods4.1 Water and water deposit samples and dilutions of thesesamples are dispensed into tubes of Starkeys medium(Aor B)following five tube MPN procedures.The tubes are sealed withliquid paraffin,and incubated at 20C for 21 days.4Positivereactions are indicated by the deposit of a black precipitate.5.Significance and Use5.1 Sulfate-reducing bacteria are widely distributed in ma-rine and fresh water muds which,in consequence,frequently1These test methods are under the jurisdiction of ASTM Committee D19 onWater and are the direct responsibility of Subcommittee D19.24 on Water Micro-biology.Current edition approved July 15,2015.Published August 2015.Originallyapproved in 1984.Last previous edition approved in 2009 as D4412 84(2009).DOI:10.1520/D4412-15.2For referenced ASTM standards,visit the ASTM website,www.astm.org,orcontact ASTM Customer Service at serviceastm.org.For Annual Book of ASTMStandards volume information,refer to the standards Document Summary page onthe ASTM website.3Available from Standard Methods,http:/standardmethods.org.4Starkey,R.L.,“Characteristics and Cultivation of Sulfate-Reducing Bacteria,”Journal of the American Water Works Association,Vol 40,1948,pp.12911298.*A Summary of Changes section appears at the end of this standardCopyright ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959.United States1 are laden with the hydrogen sulfide produced by these organ-isms during dissimilatory sulfate reduction.5.2 It has been reported that Desulfovibrio spp.can form asmuch as 10 g of sulfide per litre during active multiplication.Sulfate-reducing bacteria can cause the external or internalcorrosion of water or wastewater pipelines and pipelines forpetroleum and natural gas.The formation of galvanic cells bymassive growth of sulfate-reducing bacteria under suitableconditions makes the corrosion much worse than just the effectof the hydrogen sulfide on the metal or concrete.6.Apparatus and Materials6.1 Anaerobic Incubator,20C,if available,or conventional20C incubator.NOTE1For thermophilic organisms use a 45C incubator.6.2 Pipets,sterile,1 mL and 10 mL,“calibrated”to deliver.6.3 Test Tubes,with close fitting or airtight caps;16 by 150mm and 20 by 150 mm.6.4 Test Tube Racks,of sufficient size to contain 16 and20-mm tubes.7.Reagents7.1 Purity of ReagentsReagent grade ch