ASTM E2563-13
(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="ital">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 metal grinding 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 combined with the total mycobacterial counts (including viable culturable (VC), viable-non culturable (VNC) and non viable (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 metal working 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 and health practices and determine the applicability of regulatory limitations prior to use.
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Designation: E2563 − 13 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 3. Terminology
1.1 This practice covers the detection and enumeration of 3.1 Definitions:
viableandculturablerapidlygrowing Mycobacteria(RGM),or 3.1.1 rapidly growing mycobacteria (RGM)—non-
non-tuberculosis Mycobacteria (NTM) in aqueous metalwork- tuberculous Mycobacteria that grow and produce visible colo-
ing fluids (MWF) in the presence of high non-mycobacterial nies in four to seven days.
background population using standard microbiological culture
4. Summary of Practice
methods.
4.1 For recovery and enumeration of viable and culturable
1.2 The detection limit is one colony forming unit
Mycobacteria population in metalworking fluid field samples
(CFU)/mL metalworking fluid.
selective culture medium containing antimicrobial agents to
1.3 This practice involves culture of organisms classified as
suppress bacterial and fungal contamination is recommended.
Level 2 pathogens, and should be undertaken by a trained
(See Section 8). Standard microbiological spread and droplet
microbiologist in an appropriately equipped facility. The mi-
plating techniques are used for the enumeration of Mycobac-
crobiologist should also be capable of distinguishing the
teria. After a minimum of 14 days incubation at 30°C, the
diverse colonies of Mycobacteria from other microorganism
Mycobacteria colonies are counted and confirmed by acid-fast
colonies on a Petri dish and capable of confirming Mycobac-
staining technique specific for Mycobacteria.
teria by acid fast staining method
5. Significance and Use
1.4 This standard does not purport to address all of the
safety concerns, if any, associated with its use. It is the 5.1 Thispracticeallowsfortherecoveryandenumerationof
responsibility of the user of this standard to establish appro- viable and culturable, non-tuberculosis, rapidly growing My-
priate safety and health practices and determine the applica- cobacteria (M.immunogenum, M.chelonae, M. absessus, M.
bility of regulatory limitations prior to use.
fortuitum, and M.smegmatis) in the presence of high gram
negative background populations in metalworking fluid field
2. Referenced Documents
samples. During the past decade it has become increasingly
2
2.1 ASTM Standards: apparent that non-tuberculous Mycobacteria are common
D5465Practice for Determining Microbial Colony Counts members of the indigenous MWF bacterial population. This
from Waters Analyzed by Plating Methods population is predominantly comprised of gram negative
E1326GuideforEvaluatingNonconventionalMicrobiologi- bacteria and fungi. Mycobacterial contamination of metal-
cal Tests Used for Enumerating Bacteria working fluids has been putatively associated with hypersen-
3
2.2 Other Documents: sitivity pneumonitis (HP) amongst metal grinding machinists.
Kinyuon Acid-Fast Staining Procedure 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.
1
This practice is under the jurisdiction of ASTM Committee E34 on Occupa-
tionalHealthandSafetyandisthedirectresponsibilityofSubcommitteeE34.50on
5.2 The measurement of viable and culturable mycobacte-
Health and Safety Standards for Metal Working Fluids.
rial densities combined with the total mycobacterial counts
CurrenteditionapprovedJuly1,2013.PublishedJuly2013.Originallyapproved
(including viable culturable (VC), viable-non culturable
in 2007. Last previous edition approved in 2007 as E2563 - 07. DOI: 10.1520/
E2563-13.
(VNC) and non viable (NV) counts) is usually the first step in
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
establishing any possible relationship between Mycobacteria
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
and occupational health concerns (for example, HP).
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website.
5.3 The practice can be employed in survey studies to
3
Public Heatlth Microbiology:AGuidefortheLevelIIILaboratory.Centersfor
characterize the viable-culturable mycobacterial population
Disease Control, U.S. Department of Health and Human Services, Atlanta, GA,
1985. densities of metal working fluid field samples.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Con
...
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 − 07 E2563 − 13 An American National Standard
Standard Test Method 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 test method 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 test method 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 and health practices and determine the applicability of regulatory
limitations prior to use.
2. Referenced Documents
2
2.1 ASTM Standards:
D5465 Practice for Determining Microbial Colony Counts from Waters Analyzed by Plating Methods
E1326 Guide for Evaluating Nonconventional Microbiological Tests Used for Enumerating Bacteria
3
2.2 Other Documents:
Kinyuon Acid-Fast Staining Procedure
3. Terminology
3.1 Definitions:
3.1.1 rapidly growing mycobacteria (RGM)—non-tuberculous Mycobacteria that grow and produce visible colonies in four to
seven days.
4. Summary of Test Method
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
Section 8). Standard microbiological spread and droplet plating techniques are used for the enumeration of Mycobacteria. After
a minimum of 14 days incubation at 30°C, the Mycobacteria colonies are counted and confirmed by acid-fast staining technique
specific for Mycobacteria.
5. Significance and Use
5.1 This method 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
1
This test method 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 Oct. 1, 2007July 1, 2013. Published October 2007July 2013. Originally approved in 2007. Last previous edition approved in 2007 as E2563 -
07. DOI: 10.1520/E2563-07.10.1520/E2563-13.
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 Microbiology: A Guide for the Level III 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
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E2563 − 13
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 metal grinding 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 m
...
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