ASTM_D_5246_-_19.pdf

Designation:D524619Standard Test Method forIsolation and Enumeration of Pseudomonas aeruginosa fromWater1This standard is issued under the fixed designation D5246;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 The test method covers the isolation and enumeration ofPseudomonas aeruginosa.Testing was performed on spikedsamples using reagent grade water as the diluent from surfacewaters;recreational waters;ground water,water supplies;especially rural nonchlorinated sources;waste water;andsaline waters.The detection limit of this test method is onemicroorganism per 100 mL.1.2 This test method was used successfully with reagentwater.It is the users responsibility to ensure the validity of thistest method for surface waters,recreational waters,groundwater,rural nonchlorinated sources;waste water;and salinewaters.1.3 The values stated in SI units are to be regarded asstandard.No other units of measurement are included in thisstandard.1.4 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,health,and environmental practices and deter-mine the applicability of regulatory limitations prior to use.Specific hazard statements are given in Section 10.1.5 This international standard was developed in accor-dance with internationally recognized principles on standard-ization established in the Decision on Principles for theDevelopment of International Standards,Guides and Recom-mendations issued by the World Trade Organization TechnicalBarriers to Trade(TBT)Committee.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 WaterD3370 Practices for Sampling Water from Flowing ProcessStreams3.Terminology3.1 Definitions:3.1.1 For definitions of terms used in this standard,refer toTerminology D1129.3.2 Definitions of Terms Specific to This Standard:3.2.1 Pseudomonas aeruginosa,nan aerobic,motile,gram negative rod that produces fluorescent pigments andpyocyanin.3.2.1.1 DiscussionIt is oxidase and caseinase positive,isable to grow at 42C,is relatively resistant to many antibiotics,and may utilize acetamide.3.2.2 refrigeration,nstorage at 2 to 8C.4.Summary of Test Method4.1 A water sample is passed through a 0.45 mm orequivalent membrane filter.The filter carrying the retainedorganisms is placed on a selective medium(M-PA-C)3and isincubated at 41.5 6 0.5C for 72 h.The resulting pink-brownto black colonies of P.aeruginosa are counted and reported per100 mL of the sample.Colonies may be verified on skim milkagar.5.Significance and Use5.1 P.aeruginosa is an opportunistic pathogen and has beenlinked as the causative agent of numerous infections that maybe transmitted through a contaminated water supply to asusceptible host.NOTE1Fecal waste is 95%E.coli which is found in humans andwarm bloodied animals.5.2 The membrane filtration procedure described is a rapidand reliable test method of detecting P.aeruginosa in water.1This test method is under the jurisdiction of ASTM Committee D19 on Waterand is the direct responsibility of Subcommittee D19.24 on Water Microbiology.Current edition approved Dec.1,2019.Published December 2019.Originallyapproved in 1992.Last previous edition approved in 2015 as D5246 15.DOI:10.1520/D5246-19.2For 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 BBL Microbiological Systems,Division of Becton Dickinsonand Co.,Cockeysville,MD 21030.Other suppliers may be utilized if equivalent.*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 StatesThis international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for theDevelopment of International Standards,Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade(TBT)Committee.6.Interferences6.1 For certain samples,bacterial cells may have beenexposed to adverse environmental factors that lower theirprobability for survival and growth on a membrane filtermedium.This effect may be pronounced in this test method dueto the presence of antibiotics and the elevated incubationtemperature.6.2 The selection of an appropriate dilution volume isessential.Too small a dilution volume may fail to detect any P.ae