ASTM E2196-23
(Test Method)Standard Test Method for Quantification of Pseudomonas aeruginosa Biofilm Grown with Medium Shear and Continuous Flow Using Rotating Disk Reactor
Standard Test Method for Quantification of <emph type="bdit">Pseudomonas aeruginosa</emph > Biofilm Grown with Medium Shear and Continuous Flow Using Rotating Disk Reactor
SIGNIFICANCE AND USE
5.1 Bacteria that exist in a biofilm are phenotypically different from suspended cells of the same genotype. The study of biofilm in the laboratory requires protocols that account for this difference. Laboratory biofilms are engineered in growth reactors designed to produce a specific biofilm type. Altering system parameters will correspondingly result in a change in the biofilm. The purpose of this method is to direct a user in the laboratory study of biofilms by clearly defining each system parameter. This method will enable a person to grow, sample, and analyze a laboratory biofilm. The method was originally developed to study toilet bowl biofilms, but may also be utilized for research that requires a biofilm grown under moderate fluid shear.
SCOPE
1.1 This test method is used for growing a reproducible (1)2 Pseudomonas aeruginosa biofilm in a continuously stirred tank reactor (CSTR) under medium shear conditions. In addition, the test method describes how to sample and analyze biofilm for viable cells.
1.2 Although this test method was created to mimic conditions within a toilet bowl, it can be adapted for the growth and characterization of varying species of biofilm (rotating disk reactor—repeatability and relevance (2)).
1.3 This test method describes how to sample and analyze biofilm for viable cells. Biofilm population density is recorded as log10 colony forming units per surface area (rotating disk reactor—efficacy test method (3)).
1.4 Basic microbiology training is required to perform this test method.
1.5 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.6 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.
1.7 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.
General Information
Relations
Buy Standard
Standards Content (Sample)
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: E2196 − 23
Standard Test Method for
Quantification of Pseudomonas aeruginosa Biofilm Grown
with Medium Shear and Continuous Flow Using Rotating
1
Disk Reactor
This standard is issued under the fixed designation E2196; 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* 2. Referenced Documents
2 3
1.1 This test method is used for growing a reproducible (1) 2.1 ASTM Standards:
Pseudomonas aeruginosa biofilm in a continuously stirred tank E2756 Terminology Relating to Antimicrobial and Antiviral
reactor (CSTR) under medium shear conditions. In addition, Agents
the test method describes how to sample and analyze biofilm D5465 Practices for Determining Microbial Colony Counts
for viable cells. from Waters Analyzed by Plating Methods
2.2 Other Standards:
1.2 Although this test method was created to mimic condi-
Method 9050 C.1.a Buffered Dilution Water Preparation (4)
tions within a toilet bowl, it can be adapted for the growth and
characterization of varying species of biofilm (rotating disk
3. Terminology
reactor—repeatability and relevance (2)).
3.1 For definitions of terms used in this test method refer to
1.3 This test method describes how to sample and analyze
Terminology E2756.
biofilm for viable cells. Biofilm population density is recorded
3.2 Definitions of Terms Specific to This Standard:
as log colony forming units per surface area (rotating disk
10
3.2.1 biofilm, n—microorganisms living in a self-organized
reactor—efficacy test method (3)).
community attached to surfaces, interfaces, or each other,
1.4 Basic microbiology training is required to perform this
embedded in a matrix of extracellular polymeric substances of
test method.
microbial origin, while exhibiting altered phenotypes with
respect to growth rate and gene transcription.
1.5 The values stated in SI units are to be regarded as
3.2.1.1 Discussion—Biofilms may be comprised of bacteria,
standard. No other units of measurement are included in this
fungi, algae, protozoa, viruses, or infinite combinations of
standard.
these microorganisms. The qualitative characteristics of a
1.6 This standard does not purport to address all of the
biofilm, including, but not limited to, population density,
safety concerns, if any, associated with its use. It is the
taxonomic diversity, thickness, chemical gradients, chemical
responsibility of the user of this standard to establish appro-
composition, consistency, and other materials in the matrix that
priate safety, health, and environmental practices and deter-
are not produced by the biofilm microorganisms, are controlled
mine the applicability of regulatory limitations prior to use.
by the physiochemical environment in which it exists.
1.7 This international standard was developed in accor-
3.2.2 coupon, n—biofilm sample surface.
dance with internationally recognized principles on standard-
ization established in the Decision on Principles for the
4. Summary of Test Method
Development of International Standards, Guides and Recom-
4.1 This test method is used for growing a reproducible
mendations issued by the World Trade Organization Technical
Pseudomonas aeruginosa biofilm in a rotating disk reactor.
Barriers to Trade (TBT) Committee.
The biofilm is established by operating the reactor in batch
mode (no flow) for 24 h. Steady state growth (attachment is
equal to detachment) is reached while the reactor operates for
1
This test method is under the jurisdiction of ASTM Committee E35 on
an additional 24 h with continuous flow of the nutrients. The
Pesticides, Antimicrobials, and Alternative Control Agents and is the direct
responsibility of Subcommittee E35.15 on Antimicrobial Agents.
Current edition approved April 1, 2023. Published April 2023. Originally
3
approved in 2002. Last previous edition approved in 2022 as E2196 – 22. DOI: For referenced ASTM standards, visit the ASTM website, www.astm.org, or
10.1520/E2196-23. contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
2
The boldface numbers in parentheses refer to a list of references at the end of Standards volume information, refer to the standard’s Document Summary page on
this standard. the ASTM website.
*A Summary of Changes section appears at the end of this standard
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 ----------------------
E2196 − 23
4
NOTE 1—Al
...
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: E2196 − 22 E2196 − 23
Standard Test Method for
Quantification of Pseudomonas aeruginosa Biofilm Grown
with Medium Shear and Continuous Flow Using Rotating
1
Disk Reactor
This standard is issued under the fixed designation E2196; 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*
2
1.1 This test method is used for growing a reproducible (1) Pseudomonas aeruginosa biofilm in a continuously stirred tank
reactor (CSTR) under medium shear conditions. In addition, the test method describes how to sample and analyze biofilm for
viable cells.
1.2 Although this test method was created to mimic conditions within a toilet bowl, it can be adapted for the growth and
characterization of varying species of biofilm (rotating disk reactor—repeatability and relevance (2)).
1.3 This test method describes how to sample and analyze biofilm for viable cells. Biofilm population density is recorded as log
10
colony forming units per surface area (rotating disk reactor—efficacy test method (3)).
1.4 Basic microbiology training is required to perform this test method.
1.5 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.6 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 healthsafety, health, and environmental practices and determine
the applicability of regulatory limitations prior to use.
1.7 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
3
2.1 ASTM Standards:
E2756 Terminology Relating to Antimicrobial and Antiviral Agents
D5465 Practices for Determining Microbial Colony Counts from Waters Analyzed by Plating Methods
1
This test method is under the jurisdiction of ASTM Committee E35 on Pesticides, Antimicrobials, and Alternative Control Agents and is the direct responsibility of
Subcommittee E35.15 on Antimicrobial Agents.
Current edition approved May 1, 2022April 1, 2023. Published May 2022April 2023. Originally approved in 2002. Last previous edition approved in 20172022 as
E2196 – 17.E2196 – 22. DOI: 10.1520/E2196-22.10.1520/E2196-23.
2
The boldface numbers in parentheses refer to a list of references at the end of this standard.
3
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.
*A Summary of Changes section appears at the end of this standard
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 ----------------------
E2196 − 23
2.2 Other Standards:
Method 9050 C.1.a Buffered Dilution Water Preparation (4)
3. Terminology
3.1 For definitions of terms used in this test method refer to Terminology E2756.
3.2 Definitions of Terms Specific to This Standard:
3.2.1 biofilm, n—microorganisms living in a self-organized community attached to surfaces, interfaces, or each other, embedded
in a matrix of extracellular polymeric substances of microbial origin, while exhibiting altered phenotypes with respect to growth
rate and gene transcription.
3.2.1.1 Discussion—
Biofilms may be comprised of bacteria, fungi, algae, protozoa, viruses, or infinite combinations of these microorganisms. The
qualitative characteristics of a biofilm, including, but not limited to, population density, taxonomic diversity, thickness, chemical
gradients, chemical composition, consistency, and other materials in the matrix that are not produced by the biofilm
microorganisms, are controlled by the physiochemical environment in which it exists.
3.2.2 coupon, n—biofilm sample surface.
4. Summary of Test Method
4.1 This test method is used for growing a reproducible Pseudomonas aeruginosa biofilm in a rotating disk reactor. The biofilm
is established by operating the reactor in batch mode
...
Questions, Comments and Discussion
Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.