1、Designation:D525919Standard Test Method forIsolation and Enumeration of Enterococci from Water by theMembrane Filter Procedure1This standard is issued under the fixed designation D5259;the number immediately following the designation indicates the year oforiginal adoption or,in the case of revision,
2、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 covers a membrane filter(MF)proce-dure for the detection and enumeration of the enterococcibac
3、teria in water.The enterococci,which include Entero-coccus faecalis(E.faecalis),E.faecium,and their varieties arecommonly found in the feces of humans and other warm-blooded animals.Although some strains are ubiquitous and notrelated to fecal pollution,enterococci in water are an indicationof fecal
4、pollution and the possible presence of enteric patho-gens.These bacteria are found in water and wastewater in awide range of densities.The detection limit is one colonyforming unit(CFU)/volume filtered.1.2 This test method has been used successfully withtemperate fresh and marine ambient waters,and
5、wastewaters.Itis the users responsibility to ensure the validity of this testmethod for waters of untested types.1.3 The values stated in SI units are to be regarded asstandard.The values given in parentheses after SI units areincluded for information only and are not considered standard.1.4 This st
6、andard 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.For specific ha
7、zard statements,see Section 9.1.5 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 Orga
8、nization TechnicalBarriers to Trade(TBT)Committee.2.Referenced Documents2.1 ASTM Standards:2D1129 Terminology Relating to WaterD1193 Specification for Reagent WaterD3370 Practices for Sampling Water from Flowing ProcessStreamsD3870 Practice for Establishing Performance Characteristicsfor Colony Coun
9、ting Methods in Microbiology(With-drawn 2000)33.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 Enterococcus,nin this test method,Enterococcusspecies are those bacteria that produce red
10、 to maroon colonieswith black or reddish-brown precipitate on underside,afterincubation on mE agar and subsequent transfer to EIAmedium.3.2.1.1 DiscussionEnterococci include E.faecalis,E.faecium,E.avium,and their variants.4.Summary of Test Method4.1 The procedure given in this test method provides a
11、direct count of bacteria in water based on the development ofcolonies on the surface of the membrane filter.4Awater sampleis filtered through the membrane that retains the bacteria.Following filtration,the membrane containing the bacterialcells is placed on a selective,medium,mE agar,and incubated1T
12、his 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 April 1,2019.Published April 2019.Originallyapproved in 1992.Last previous edition approved in 2014 as D5259 14.DOI:10.1520/D
13、5259-19.2For 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.3The last approved version of this historical standard is
14、referenced onwww.astm.org.4Cabelli,V.J.,Dufour,A.P.,Levin,M.A.,McCabe,L.J.,and Haberman,P.W.,“Relationship of Microbial Indicators to Health Effects at Marine Bathing Beaches,”American Journal of Public Health,Vol 69,1979,pp.690696.Copyright ASTM International,100 Barr Harbor Drive,PO Box C700,West
15、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 O
16、rganization Technical Barriers to Trade(TBT)Committee.1 for 48 h at 41C,then transferred to EIA agar and held at 41Cfor 20 min.Enterococci develop as red to maroon colonies withblack or reddish-brown precipitate on the underside of thefilter.5.Significance and Use5.1 The enterococci are indicators of the bacteriologicalquality for potable water,shellfish growing waters,ambient,and recreational waters.A direct relationship betweenswimming,associated gastroenteritis,and enterococci has beenestabli
