ASTM_D_8429_-_21.pdf

Designation:D842921Standard Test Method forLegionella pneumophila in Water Samples Using Legiolert1,2This standard is issued under the fixed designation D8429;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.Scope1.1 This test method describes a simple procedure for thedetection of Legionella pneumophila(L.pneumophila)inpotable water and non-potable waters(cooling towers,forexample).This procedure describes a liquid culture methodbased on a bacterial enzyme technology.The detection of L.pneumophila is signaled through the utilization of a substratepresent in the Legiolert reagent.L.pneumophila cells growrapidly and reproduce using the rich supply of amino acids,vitamins and other nutrients present in the Legiolert reagent.Actively growing strains of L.pneumophila use the addedsubstrate to produce a brown color indicator or produce turbidgrowth with or without brown coloration.Legiolert can detectthis bacterial species at the following minimum concentrationsbased on the protocol employed:1.1.1 Potable Water:1.1.1.1 1 organism/100 mL at 7 days for 100 mL potableprotocol.1.1.1.2 1 organism/10 mL at 7 days for 10 mL potableprotocol.1.1.2 Non-potable Water:1.1.2.1 1 organism/1.0 mL at 7 days for 1.0 mLnon-potable protocol.1.1.2.2 1 organism/0.1 mL at 7 days for 0.1 mLnon-potable protocol.1.1.3 This test method can be used for potable(drinking)waters and non-potable waters such as cooling tower waters(1).3It is the users responsibility to ensure the validity of thistest method for waters of untested matrices.1.2 The values stated in SI units are to be regarded asstandard.No other units of measurement are included in thisstandard.1.3 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.1.4 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:4D1129 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 ProcessStreamsD5952 Guide for the Inspection of Water Systems forLegionella and the Investigation of Possible Outbreaks ofLegionellosis(Legionnaires Disease or Pontiac Fever)D6503 Test Method for Enterococci in Water Using Enter-olert3.Terminology3.1 Definitions:3.1.1 For definitions of terms used in this standard,refer toTerminology D1129.3.1.2 Legionella,na bacterial genus containing over 50species and at least 71 serogroups;abbreviated to the firstinitial when used relatedly with a species name,for example,L.pneumophila.3.1.3 most probable number(MPN),na statistical methodfor determining bacterial concentration based on the Poissondistribution.3.2 Definitions of Terms Specific to This Standard:3.2.1 Legionella pneumophila,nin the context of thismethod,a pathogenic gram-negative bacteria that produces anydegree of brown coloration or turbid growth with or withoutbrown coloration in the presence of the Legiolert reagent.1Legiolert is a trademark of IDEXX Laboratories,Inc.,Westbrook,ME.2This 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.1,2021.Published December 2021.DOI:10.1520/D8429-21.3The boldface numbers in parentheses refer to a list of references at the end ofthis standard.4For 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.Copyright 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.1?3.2.2 Quanti-Tray/Legiolert,na system for the quantifica-tion of Legionella pneumophila.It consists of a Quanti-TraySealer PLUS5,6and 96-multi well Quanti-Trays that express L.pneumophila counts as MPN per volume of sample.3.2.2.1 Di