1、Designation:E256318An American National StandardStandard Practice forEnumeration of Non-Tuberculosis Mycobacteria in AqueousMetalworking Fluids by Plate Count Method1This standard is issued under the fixed designation E2563;the number immediately following the designation indicates the year oforigin
2、al 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 practice covers the detection and enumeration ofviable and culturab
3、le rapidly growing Mycobacteria(RGM),ornon-tuberculosis Mycobacteria(NTM)in aqueous metalwork-ing fluids(MWF)in the presence of high non-mycobacterialbackground population using standard microbiological culturemethods.1.2 The detection limit is one colony forming unit(CFU)/mL metalworking fluid.1.3
4、This practice involves culture of organisms classified asLevel 2 pathogens,and should be undertaken by a trainedmicrobiologist in an appropriately equipped facility.The mi-crobiologist should also be capable of distinguishing thediverse colonies of Mycobacteria from other microorganismcolonies on a
5、Petri dish and capable of confirming Mycobac-teria by acid-fast staining method.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 practi
6、ces and deter-mine the applicability of regulatory limitations prior to use.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 R
7、ecom-mendations issued by the World Trade Organization TechnicalBarriers to Trade(TBT)Committee.2.Referenced Documents2.1 ASTM Standards:2D5465 Practices for Determining Microbial Colony Countsfrom Waters Analyzed by Plating MethodsE1326 Guide for Evaluating Non-culture MicrobiologicalTestsE2523 Ter
8、minology for Metalworking Fluids and Opera-tions2.2 Other Document:3Kinyoun Acid-Fast Staining Procedure3.Terminology3.1 For definitions of terms used in this standard,refer toTerminology E2523.3.2 Definitions:3.2.1 rapidlygrowingmycobacteria(RGM)non-tuberculous Mycobacteria that grow and produce vi
9、sible colo-nies in four to seven days.4.Summary of Practice4.1 For recovery and enumeration of viable and culturableMycobacteria population in metalworking fluid field samples,selective culture medium containing antimicrobial agents tosuppress bacterial and fungal contamination is recommended.(See S
10、ection 8.)Standard microbiological spread and dropletplating techniques are used for the enumeration of Mycobac-teria(see Practices D5465).After a minimum of 14 daysincubation at 30 C,the Mycobacteria colonies are countedand confirmed by acid-fast staining technique specific forMycobacteria.5.Signif
11、icance and Use5.1 This practice allows for the recovery and enumeration ofviable and culturable,non-tuberculosis,rapidly growing My-cobacteria(M.immunogenum,M.chelonae,M.absessus,M.fortuitum,and M.smegmatis)in the presence of high Gram-negative background populations in metalworking fluid fieldsampl
12、es.During the past decade,it has become increasinglyapparent that non-tuberculous Mycobacteria are commonmembers of the indigenous MWF bacterial population.Thispopulation is predominantly comprised of Gram-negative bac-teria and fungi.Mycobacterial contamination of metalworking1This practice is unde
13、r the jurisdiction of ASTM Committee E34 on Occupa-tional Health and Safety and is the direct responsibility of Subcommittee E34.50 onHealth and Safety Standards for Metal Working Fluids.Current edition approved Oct.1,2018.Published October 2018.Originallyapproved in 2007.Last previous edition appro
14、ved in 2013 as E2563 13.DOI:10.1520/E2563-18.2For 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.3Public Health Microb
15、iology:A Guide for the Level III Laboratory,Centers forDisease Control,U.S.Department of Health and Human Services,Atlanta,GA,1985.Copyright ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959.United StatesThis international standard was developed in accordance with
16、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 fluids has been putatively associated with hypersensitivitypneumonitis(HP)amongst metalgrinding machinists.Thedetection and enumeration of these organisms will aid in betterunderstanding of occupational health-related problems and abetter assess
