ASTM E2563-18
(Practice)Standard Practice for Enumeration of Non-Tuberculosis Mycobacteria in Aqueous Metalworking Fluids by Plate Count Method
Standard Practice for Enumeration of Non-Tuberculosis <emph type="bdit">Mycobacteria</emph > in Aqueous Metalworking Fluids by Plate Count Method
SIGNIFICANCE AND USE
5.1 This practice allows for the recovery and enumeration of viable and culturable, non-tuberculosis, rapidly growing Mycobacteria (M. immunogenum, M. chelonae, M. absessus, M. fortuitum, and M. smegmatis) in the presence of high Gram-negative background populations in metalworking fluid field samples. During the past decade, it has become increasingly apparent that non-tuberculous Mycobacteria are common members of the indigenous MWF bacterial population. This population is predominantly comprised of Gram-negative bacteria and fungi. Mycobacterial contamination of metalworking fluids has been putatively associated with hypersensitivity pneumonitis (HP) amongst metalgrinding machinists. The detection and enumeration of these organisms will aid in better understanding of occupational health-related problems and a better assessment of antimicrobial pesticide efficacy.
5.2 The measurement of viable and culturable mycobacterial densities (Guide E1326), combined with the total mycobacterial counts (including viable culturable (VC), viable nonculturable (VNC) and nonviable (NV) counts), is usually the first step in establishing any possible relationship between Mycobacteria and occupational health concerns (for example, HP).
5.3 The practice can be employed in survey studies to characterize the viable-culturable mycobacterial population densities of metalworking fluid field samples.
5.4 This practice is also applicable for establishing the mycobacterial resistance of metalworking fluid formulations by determining mycobacterium survival by means of plate count technique.
5.5 This practice can also be used to evaluate the relative efficacy of microbicides against Mycobacteria in metalworking fluids.
SCOPE
1.1 This practice covers the detection and enumeration of viable and culturable rapidly growing Mycobacteria (RGM), or non-tuberculosis Mycobacteria (NTM) in aqueous metalworking fluids (MWF) in the presence of high non-mycobacterial background population using standard microbiological culture methods.
1.2 The detection limit is one colony forming unit (CFU)/mL metalworking fluid.
1.3 This practice involves culture of organisms classified as Level 2 pathogens, and should be undertaken by a trained microbiologist in an appropriately equipped facility. The microbiologist should also be capable of distinguishing the diverse colonies of Mycobacteria from other microorganism colonies on a Petri dish and capable of confirming Mycobacteria by acid-fast staining method.
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.
1.5 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
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This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation: E2563 − 18 An American National Standard
Standard Practice for
Enumeration of Non-Tuberculosis Mycobacteria in Aqueous
1
Metalworking Fluids by Plate Count Method
This standard is issued under the fixed designation E2563; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope E1326Guide for Evaluating Non-culture Microbiological
Tests
1.1 This practice covers the detection and enumeration of
E2523Terminology for Metalworking Fluids and Opera-
viableandculturablerapidlygrowing Mycobacteria(RGM),or
tions
non-tuberculosis Mycobacteria (NTM) in aqueous metalwork-
3
2.2 Other Document:
ing fluids (MWF) in the presence of high non-mycobacterial
Kinyoun Acid-Fast Staining Procedure
background population using standard microbiological culture
methods.
3. Terminology
1.2 The detection limit is one colony forming unit
3.1 For definitions of terms used in this standard, refer to
(CFU)/mL metalworking fluid.
Terminology E2523.
1.3 This practice involves culture of organisms classified as
3.2 Definitions:
Level 2 pathogens, and should be undertaken by a trained
3.2.1 rapidly growing mycobacteria (RGM)—non-
microbiologist in an appropriately equipped facility. The mi-
tuberculous Mycobacteria that grow and produce visible colo-
crobiologist should also be capable of distinguishing the
nies in four to seven days.
diverse colonies of Mycobacteria from other microorganism
colonies on a Petri dish and capable of confirming Mycobac-
4. Summary of Practice
teria by acid-fast staining method.
4.1 For recovery and enumeration of viable and culturable
1.4 This standard does not purport to address all of the
Mycobacteria population in metalworking fluid field samples,
safety concerns, if any, associated with its use. It is the
selective culture medium containing antimicrobial agents to
responsibility of the user of this standard to establish appro-
suppress bacterial and fungal contamination is recommended.
priate safety, health, and environmental practices and deter-
(See Section 8.) Standard microbiological spread and droplet
mine the applicability of regulatory limitations prior to use.
plating techniques are used for the enumeration of Mycobac-
1.5 This international standard was developed in accor-
teria (see Practices D5465). After a minimum of 14 days
dance with internationally recognized principles on standard-
incubation at 30°C, the Mycobacteria colonies are counted
ization established in the Decision on Principles for the
and confirmed by acid-fast staining technique specific for
Development of International Standards, Guides and Recom-
Mycobacteria.
mendations issued by the World Trade Organization Technical
Barriers to Trade (TBT) Committee.
5. Significance and Use
5.1 Thispracticeallowsfortherecoveryandenumerationof
2. Referenced Documents
viable and culturable, non-tuberculosis, rapidly growing My-
2
2.1 ASTM Standards:
cobacteria (M. immunogenum, M. chelonae, M. absessus, M.
D5465Practices for Determining Microbial Colony Counts
fortuitum, and M. smegmatis) in the presence of high Gram-
from Waters Analyzed by Plating Methods
negative background populations in metalworking fluid field
samples. During the past decade, it has become increasingly
1
apparent that non-tuberculous Mycobacteria are common
This practice is under the jurisdiction of ASTM Committee E34 on Occupa-
tionalHealthandSafetyandisthedirectresponsibilityofSubcommitteeE34.50on
members of the indigenous MWF bacterial population. This
Health and Safety Standards for Metal Working Fluids.
population is predominantly comprised of Gram-negative bac-
Current edition approved Oct. 1, 2018. Published October 2018. Originally
teria and fungi. Mycobacterial contamination of metalworking
approved in 2007. Last previous edition approved in 2013 as E2563 – 13. DOI:
10.1520/E2563-18.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
3
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM Public Health Microbiology: A Guide for the Level III Laboratory, Centers for
Standards volume information, refer to the standard’s Document Summary page on Disease Control, U.S. Department of Health and Human Services, Atlanta, GA,
the ASTM website. 1985.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 ----------------------
E2563 − 18
fluids has been putatively associated
...
This document is not an ASTM standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Because
it may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current version
of the standard as published by ASTM is to be considered the official document.
Designation: E2563 − 13 E2563 − 18 An American National Standard
Standard Practice for
Enumeration of Non-Tuberculosis Mycobacteria in Aqueous
1
Metalworking Fluids by Plate Count Method
This standard is issued under the fixed designation E2563; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope
1.1 This practice covers the detection and enumeration of viable and culturable rapidly growing Mycobacteria (RGM), or
non-tuberculosis Mycobacteria (NTM) in aqueous metalworking fluids (MWF) in the presence of high non-mycobacterial
background population using standard microbiological culture methods.
1.2 The detection limit is one colony forming unit (CFU)/mL metalworking fluid.
1.3 This practice involves culture of organisms classified as Level 2 pathogens, and should be undertaken by a trained
microbiologist in an appropriately equipped facility. The microbiologist should also be capable of distinguishing the diverse
colonies of Mycobacteria from other microorganism colonies on a Petri dish and capable of confirming Mycobacteria by acid fast
acid-fast staining methodmethod.
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility
of the user of this standard to establish appropriate safety safety, health, and healthenvironmental practices and determine the
applicability of regulatory limitations prior to use.
1.5 This international standard was developed in accordance with internationally recognized principles on standardization
established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued
by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
2. Referenced Documents
2
2.1 ASTM Standards:
D5465 Practices for Determining Microbial Colony Counts from Waters Analyzed by Plating Methods
E1326 Guide for Evaluating Non-culture Microbiological Tests
E2523 Terminology for Metalworking Fluids and Operations
3
2.2 Other Documents:Document:
KinyuonKinyoun Acid-Fast Staining Procedure
3. Terminology
3.1 For definitions of terms used in this standard, refer to Terminology E2523.
3.2 Definitions:
3.2.1 rapidly growing mycobacteria (RGM)—non-tuberculous Mycobacteria that grow and produce visible colonies in four to
seven days.
4. Summary of Practice
4.1 For recovery and enumeration of viable and culturable Mycobacteria population in metalworking fluid field samples,
selective culture medium containing antimicrobial agents to suppress bacterial and fungal contamination is recommended. (See
1
This practice is under the jurisdiction of ASTM Committee E34 on Occupational Health and Safety and is the direct responsibility of Subcommittee E34.50 on Health
and Safety Standards for Metal Working Fluids.
Current edition approved July 1, 2013Oct. 1, 2018. Published July 2013October 2018. Originally approved in 2007. Last previous edition approved in 20072013 as E2563
- 07.– 13. DOI: 10.1520/E2563-13.10.1520/E2563-18.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM Standards
volume information, refer to the standard’s Document Summary page on the ASTM website.
3
Public Heatlth MicrobiologyHealth Microbiology:: A Guide for the Level III Laboratory.Laboratory, Centers for Disease 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 States
1
---------------------- Page: 1 ----------------------
E2563 − 18
Section 8).) Standard microbiological spread and droplet plating techniques are used for the enumeration of Mycobacteria.My-
cobacteria (see Practices D5465). After a minimum of 14 days incubation at 30°C,30 °C, the Mycobacteria colonies are counted
and confirmed by acid-fast staining technique specific for Mycobacteria.
5. Significance and Use
5.1 This practice allows for the recovery and enumeration of viable and culturable, non-tuberculosis, rapidly growing
Mycobacteria (M.immunogenum,M. immunogenum,M.chelonae,M. chelonae,M. absessus,M. fortuitum, and M.smegmatisM.
smegmatis) in the presence of high gram negative Gram-negative background populations in metalworking fluid fi
...
NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation: E2563 − 18 An American National Standard
Standard Practice for
Enumeration of Non-Tuberculosis Mycobacteria in Aqueous
1
Metalworking Fluids by Plate Count Method
This standard is issued under the fixed designation E2563; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope E1326 Guide for Evaluating Non-culture Microbiological
Tests
1.1 This practice covers the detection and enumeration of
E2523 Terminology for Metalworking Fluids and Opera-
viable and culturable rapidly growing Mycobacteria (RGM), or
tions
non-tuberculosis Mycobacteria (NTM) in aqueous metalwork-
3
2.2 Other Document:
ing fluids (MWF) in the presence of high non-mycobacterial
Kinyoun Acid-Fast Staining Procedure
background population using standard microbiological culture
methods.
3. Terminology
1.2 The detection limit is one colony forming unit
3.1 For definitions of terms used in this standard, refer to
(CFU)/mL metalworking fluid.
Terminology E2523.
1.3 This practice involves culture of organisms classified as
3.2 Definitions:
Level 2 pathogens, and should be undertaken by a trained
3.2.1 rapidly growing mycobacteria (RGM)—non-
microbiologist in an appropriately equipped facility. The mi-
tuberculous Mycobacteria that grow and produce visible colo-
crobiologist should also be capable of distinguishing the
nies in four to seven days.
diverse colonies of Mycobacteria from other microorganism
colonies on a Petri dish and capable of confirming Mycobac-
4. Summary of Practice
teria by acid-fast staining method.
4.1 For recovery and enumeration of viable and culturable
1.4 This standard does not purport to address all of the
Mycobacteria population in metalworking fluid field samples,
safety concerns, if any, associated with its use. It is the
selective culture medium containing antimicrobial agents to
responsibility of the user of this standard to establish appro-
suppress bacterial and fungal contamination is recommended.
priate safety, health, and environmental practices and deter-
(See Section 8.) Standard microbiological spread and droplet
mine the applicability of regulatory limitations prior to use.
plating techniques are used for the enumeration of Mycobac-
1.5 This international standard was developed in accor-
teria (see Practices D5465). After a minimum of 14 days
dance with internationally recognized principles on standard-
incubation at 30 °C, the Mycobacteria colonies are counted
ization established in the Decision on Principles for the
and confirmed by acid-fast staining technique specific for
Development of International Standards, Guides and Recom-
Mycobacteria.
mendations issued by the World Trade Organization Technical
Barriers to Trade (TBT) Committee.
5. Significance and Use
5.1 This practice allows for the recovery and enumeration of
2. Referenced Documents
viable and culturable, non-tuberculosis, rapidly growing My-
2
2.1 ASTM Standards:
cobacteria (M. immunogenum, M. chelonae, M. absessus, M.
D5465 Practices for Determining Microbial Colony Counts
fortuitum, and M. smegmatis) in the presence of high Gram-
from Waters Analyzed by Plating Methods
negative background populations in metalworking fluid field
samples. During the past decade, it has become increasingly
1
apparent that non-tuberculous Mycobacteria are common
This practice is under the jurisdiction of ASTM Committee E34 on Occupa-
tional Health and Safety and is the direct responsibility of Subcommittee E34.50 on
members of the indigenous MWF bacterial population. This
Health and Safety Standards for Metal Working Fluids.
population is predominantly comprised of Gram-negative bac-
Current edition approved Oct. 1, 2018. Published October 2018. Originally
teria and fungi. Mycobacterial contamination of metalworking
approved in 2007. Last previous edition approved in 2013 as E2563 – 13. DOI:
10.1520/E2563-18.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
3
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM Public Health Microbiology: A Guide for the Level III Laboratory, Centers for
Standards volume information, refer to the standard’s Document Summary page on Disease Control, U.S. Department of Health and Human Services, Atlanta, GA,
the ASTM website. 1985.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 ----------------------
E2563 − 18
fluids has been putatively associated with hypersensitivity 8.5 Sterile 1 mL Pipets, with 0.01 mL divisions.
pneumonitis (HP) amongst metalgrindin
...
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