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ASTM E2563-07

(Test Method)

Standard Test Method for Enumeration of Non-Tuberculosis Mycobacteria in Aqueous Metalworking Fluids by Plate Count Method

Standard Test Method for Enumeration of Non-Tuberculosis Mycobacteria in Aqueous Metalworking Fluids by Plate Count Method

SIGNIFICANCE AND USE
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 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.
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).
The method can be employed in survey studies to characterize the viable-culturable mycobacterial population densities of metal working fluid field samples.
This method is also applicable for establishing the mycobacterial resistance of metalworking fluid formulations by determining mycobacterium survival by means of plate count technique.
This method can also be used to evaluate the relative efficacy of microbicides against Mycobacteria in metalworking fluids.
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
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.

General Information

Status
Historical
Publication Date
30-Sep-2007
ICS
75.100 - Lubricants, industrial oils and related products
Technical Committee
E34 - Occupational Health and Safety
Drafting Committee
E34.50 - Health and Safety Standards for Metal Working Fluids
Current Stage
Ref Project

Relations

Replaced By
ASTM E2563-13 - Standard Practice for Enumeration of Non-Tuberculosis <emph type="ital">Mycobacteria</emph > in Aqueous Metalworking Fluids by Plate Count Method
Effective Date
01-Jul-2013
Referred By
ASTM E2889-12(2017) - Standard Practice for Control of Respiratory Hazards in the Metal Removal Fluid Environment
Effective Date
01-Oct-2007
Referred By
ASTM E2275-19 - Standard Practice for Evaluating Water-Miscible Metalworking Fluid Bioresistance and Antimicrobial Pesticide Performance
Effective Date
01-Oct-2007
Referred By
ASTM E2523-13(2018) - Standard Terminology for Metalworking Fluids and Operations
Effective Date
01-Oct-2007
Referred By
ASTM D5465-16(2020) - Standard Practices for Determining Microbial Colony Counts from Waters Analyzed by Plating Methods
Effective Date
01-Oct-2007
Referred By
ASTM E2148-21 - Standard Guide for Using Documents Related to Metalworking or Metal Removal Fluid Health and Safety
Effective Date
01-Oct-2007

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ASTM E2563-07 - Standard Test Method for Enumeration of Non-Tuberculosis Mycobacteria in Aqueous Metalworking Fluids by Plate Count MethodASTM E2563-07 - Standard Test Method for Enumeration of Non-Tuberculosis Mycobacteria in Aqueous Metalworking Fluids by Plate Count Method
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ASTM E2563-07 - Standard Test Method for Enumeration of Non-Tuberculosis Mycobacteria in Aqueous Metalworking Fluids by Plate Count Method
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Standards Content (Sample)

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 − 07 AnAmerican National Standard
Standard Test Method 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 test method covers the detection and enumeration 3.1 Definitions:
ofviableandculturablerapidlygrowing Mycobacteria(RGM), 3.1.1 rapidly growing mycobacteria (RGM)—non-
or non-tuberculosis Mycobacteria (NTM) in aqueous metal- tuberculous Mycobacteria that grow and produce visible colo-
working fluids (MWF) in the presence of high non- nies in four to seven days.
mycobacterial background population using standard micro-
4. Summary of Test Method
biological culture 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 test method involves culture of organisms classi-
suppress bacterial and fungal contamination is recommended.
fied as Level 2 pathogens, and should be undertaken by a
(See Section 8). Standard microbiological spread and droplet
trained microbiologist in an appropriately equipped facility.
plating techniques are used for the enumeration of Mycobac-
Themicrobiologistshouldalsobecapableofdistinguishingthe
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 Thismethodallowsfortherecoveryandenumerationof
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
apparent that non-tuberculous Mycobacteria are common
2.1 ASTM Standards:
members of the indigenous MWF bacterial population. This
D5465Practice for Determining Microbial Colony Counts
population is predominantly comprised of gram negative
from Waters Analyzed by Plating Methods
bacteria and fungi. Mycobacterial contamination of metal-
E1326GuideforEvaluatingNonconventionalMicrobiologi-
working fluids has been putatively associated with hypersen-
cal Tests Used for Enumerating Bacteria
3
sitivity pneumonitis (HP) amongst metal grinding machinists.
2.2 Other Documents:
The detection and enumeration of these organisms will aid in
Kinyuon Acid-Fast Staining Procedure
better understanding of occupational health related problems
and a better assessment of antimicrobial pesticide efficacy.
1
This test method is under the jurisdiction of ASTM Committee E34 on
5.2 The measurement of viable and culturable mycobacte-
Occupational Health and Safety and is the direct responsibility of Subcommittee
rial densities combined with the total mycobacterial counts
E34.50 on Health and Safety Standards for Metal Working Fluids.
(including viable culturable (VC), viable-non culturable
Current edition approved Oct. 1, 2007. Published October 2007. DOI: 10.1520/
E2563-07.
(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 method 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 Conshohocken, PA 19428-2959. United States
1

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3.1 Personnel from a wide range of disciplines contribute to metalworking fluid management and plant environment health and safety management. Consequently, terms familiar to some stakeholders will be unfamiliar to others.  
3.2 This terminology standard provides, in a single document, a compilation of definitions used by personnel involved with both metalworking environment health and safety and fluid management.  
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1.1 This terminology standard provides a compilation of ASTM and non-ASTM consensus definitions of terms used in the metalworking industry.  
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5.1 Measurement of mycobacterial cell count densities is an important step in establishing a possible relationship between mycobacteria and occupational health-related allergic responses, for example, hypersensitivity pneumonitis (HP) in persons exposed to aerosols of metalworking fluids. It is known that the viable mycobacteria count underestimates the total mycobacterial levels by not counting the non-culturable, possibly dead or moribund population that is potentially equally important in the investigation of occupational health-related problems. The direct microscopic counting method (DMC) described here gives a quantitative assessment of the total numbers of acid-fast bacilli. It involves using acid-fast staining to selectively identify mycobacteria from other bacteria, followed by enumeration or direct microscopic counting of a known volume over a known area. Although other microbes—particularly the Actinomycetes—also stain acid-fast, they are differentiated from the mycobacteria because of their morphology and size. Non-mycobacteria, acid-fast microbes are 50 to 100 times larger than mycobacteria. This practice provides quantitative information on the total (culturable and non-culturable viable, and non-viable) mycobacteria populations. The results are expressed quantitatively as mycobacteria per mL of metalworking fluid sample.  
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1.1 This practice describes a direct microscopic counting method (DMC) for the enumeration of the acid-fast stained mycobacteria population in metalworking fluids. It can be used to detect levels of total mycobacteria population, including culturable as well as non-culturable (possibly dead or moribund) bacterial cells. This practice is recommended for all water-based metalworking fluids (Classification D2881).  
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1.3 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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ASTM E2563-23(Practice)
Standard Practice for Enumeration of Non-Tuberculosis Mycobacteria 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. 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.  
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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.  
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4.1 Use of this practice will improve management and control of metal removal fluids. The proper management and use will reduce dermal and other occupational hazards associated with these fluids.  
4.1.1 Guide E2148 covers information on how to use documents related to health and safety of metalworking and metal removal fluids, including this document. Documents referenced in Guide E2148 are grouped as applicable to producers, to users, or to all.  
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1.3 Straight oils generally consist of a severely solvent-refined or hydro-treated petroleum oil, a synthetic oil, or other oils of animal or vegetable origin, including oils that are modified for performance characteristics (for example, esterified rapeseed oil, and so forth). Straight oils are not intended to be diluted with water prior to use. Additives are often included in straight oil formulations.  
1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
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5.1 The determination of endotoxin concentrations in MWF is a parameter that can be used in decision-making for prudent fluid management practices (fluid draining, cleaning, recharging, or biocide dosages).  
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5.1 This method measures the concentration of ATP present in the sample. ATP is a constituent of all living cells, including bacteria and fungi. Consequently, the presence of ATP is an indicator of total microbial contamination in metalworking fluids. ATP is not associated with matter of non-biological origin.  
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5.4 The ATP test provides rapid test results that reflect the total bioburden in the sample. It thereby reduces the delay between test initiation and data capture, from the 36 h to 48 h (or longer) required for culturable colonies to become visible, to approximately 5 min.  
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5.5.2.2 Within a species, cells that are more metabolically active will have more ATP per cell than dormant cells.
5.5.2.3 The greater the total bioburden, the greater the ATP concentration in a sample.  
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5.5.3...
SCOPE
1.1 This test method provides a protocol for capturing, extracting, and quantifying the adenosine triphosphate (ATP) content associated with microorganisms found in water-miscible metalworking fluids (MWFs).  
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1.3 This test method is equally suitable for use in the laboratory or field.  
1.4 The test method detects ATP concentrations in the range of 4.0 pg ATP/mL to 400 000 pg ATP/mL.  
1.5 Providing interferences can be overcome, bioluminescence is a reliable and proven method for qualifying and quantifying ATP. The method does not differentiate between ATP from different sources, for example, from different types of microorganisms, such as bacteria and fungi.  
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SIGNIFICANCE AND USE
4.1 The process of recirculating MWFs entrains air bubbles which can accumulate, forming foam.  
4.2 Optimally, air bubbles burst open quickly after they are created. However, air bubble persistence is affected by MWF chemistry and the mechanisms by which energy is introduced into recirculating MWFs.  
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4.3.3 Foul filters;  
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4.3.7 Contribute to safety and hygiene hazards in the plant.  
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SCOPE
1.1 This guide provides an overview of foaming tendency evaluation protocols and their appropriate use.  
1.2 ASTM Test Methods D3519 and D3601 were withdrawn in 2013. Although each method had some utility, neither method reliably predicted in-use foaming tendency. Since Test Methods D3519 and D3601 were first adopted, several more predictive test protocols have been developed. However, it is also common knowledge that no single protocol is universally suitable for predicting water-miscible metalworking fluid (MWF) foaming tendency.  
1.3 Moreover, there are no generally recognized reference standard fluids (either MWF or foam-control additive). Instead it is important to include a relevant reference sample in all testing.  
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1.5 The dilution water quality can have a major impact on foaming properties. In general, fluid concentrates diluted with hard water will foam less than those diluted with soft, deionized, or reverse osmosis water. Screening tests using the expected range of dilution water quality are highly recommended.  
1.6 The temperature of the tested fluids can have a major impact on foaming properties. In general, test fluids should be held and tested at temperatures that closely mimic the real-world application and process.  
1.7 Cleanliness of test apparatus is critical during foam evaluation testing. Traces of residue on labware can significantly impact the observed foaming tendency of a test fluid. Best practice is to clean any glassware or other vessels using some version of a chemical cleaner that will alleviate any risk of cross contamination.  
1.8 Units—The values stated in SI units are to be regarded as the standard. No other units of measurement are included in this standard.  
1.9 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 sa...

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ASTM E1687-19(Test Method)
Standard Test Method for Determining Carcinogenic Potential of Virgin Base Oils in Metalworking Fluids

SIGNIFICANCE AND USE
5.1 The test method is based on a modification of the Ames Salmonella mutagenesis assay. As modified, there is good correlation with mouse skin-painting bioassay results for samples of raw and refined lubricating oil process streams.  
5.2 Mutagenic potency in this modified assay and carcinogenicity in the skin-painting bioassay also correlate with the content of three to seven-ring PACs, which include polycyclic aromatic hydrocarbons and their heterocyclic analogs. The strength of these correlations implies that PACs are the principal mutagenic and carcinogenic species in these oils. Some of the methods that have provided evidence supporting this view are referenced in Appendix X1.
SCOPE
1.1 This test method covers a microbiological test procedure based upon the Salmonella mutagenesis assay of Ames et al. (1)2 (see also Maron et al. (2)). It can be used as a screening technique to detect the presence of potential dermal carcinogens in virgin base oils used in the formulation of metalworking oils. Persons who perform this test should be well versed in the conduct of the Ames test and conversant with the physical and chemical properties of petroleum products.  
1.2 The test method is not recommended as the sole testing procedure for oils which have viscosities less than 18 cSt (90 SUS) at 40 °C, or for formulated metalworking fluids.  
1.3 The values stated in SI units are to be regarded as the standard. The values given in parentheses are provided for information only.  
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. Section 7 provides general guidelines for safe conduct of this test method.  
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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Standard Practice for Prevention of Dermatitis in the Wet Metal Removal Fluid Environment

SIGNIFICANCE AND USE
5.1 Use of this practice is intended to reduce occupational dermatitis caused by exposure to the wet metal removal environment.  
5.2 Complaints of dermatitis conditions are often associated with exposures to metal removal fluid.  
5.3 Implementation of this practice and incorporation of metal removal fluid management program has the potential to reduce complaints of occupational dermatitis. Elements of an effective program include: understanding dermatitis and associated causes; prevention of dermatitis and exposure to metal removal fluids; appropriate product selection; good management of additives, microorganisms, and fluids; appropriate additive (including antimicrobial pesticides) selection and additive control; appropriate tool design and assessment; and control of metal removal fluid exposures, including aerosols.
SCOPE
1.1 This practice sets forth guidelines for reducing dermatitis caused by exposure to the wet metal removal environment. The scope of this practice does not include exposure to chemicals that enter the body through intact skin (cutaneous route), which has the potential to cause other toxic effects.  
1.2 This practice incorporates means and mechanisms to reduce dermal exposure to the wet metal removal environment and to control factors in the wet metal removal environment that have the potential to cause dermatitis.  
1.3 This practice focuses on employee exposure to the skin via contact and exposure to metal removal fluid (MRF).  
1.4 The values stated in inch-pound units are to be regarded as standard. The values given in parentheses are mathematical conversions to SI units that are provided for information only and are not considered standard.  
1.5 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.  
1.6 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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ASTM
ASTM D5707-23(Test Method)
Standard Test Method for Measuring Friction and Wear Properties of Lubricating Grease Using a High-Frequency, Linear-Oscillation (SRV) Test Machine

SIGNIFICANCE AND USE
5.1 This test method can be used to determine wear properties and coefficient of friction of lubricating greases at selected temperatures and loads specified for use in applications where high-speed vibrational or start-stop motions are present for extended periods of time under initial high Hertzian point contact pressures. This test method has found application in qualifying lubricating greases used in constant velocity joints of front-wheel-drive automobiles and for lubricating greases used in roller bearings. Users of this test method should determine whether results correlate with field performance or other applications.
SCOPE
1.1 This test method covers a procedure for determining a lubricating grease's coefficient of friction and its ability to protect against wear when subjected to high-frequency, linear-oscillation motion using an SRV test machine at a test load of 200 N, frequency of 50 Hz, stroke amplitude of 1.00 mm, duration of 2 h, and temperature within the range of the test machine, specifically, ambient to 280 °C. Other test loads (10 N to 1200 N for SRVI-model, 10 N to 1400 N for SRVII-model, and 10 N to 2000 N for SRVIII-model), frequencies (5 Hz to 500 Hz) and stroke amplitudes (0.1 mm up to 4.0 mm) can be used, if specified. The precision of this test method is based on the stated parameters and test temperatures of 50 °C and 80 °C. Average wear scar dimensions on ball and coefficient of friction are determined and reported.
Note 1: Optimol Instruments supplies an upgrade kit to allow SRVI/II-machines to operate with 1600 N, if needed.  
1.2 This test method can also be used for determining a fluid lubricant's ability to protect against wear and its coefficient of friction under similar test conditions.  
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
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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