ASTM E2871-21

Standard Test Method for Determining Disinfectant Efficacy Against Biofilm Grown in the CDC Biofilm Reactor Using the Single Tube Method

SIGNIFICANCE AND USE
5.1 The Single Tube Method is designed to evaluate the efficacy of disinfectants against biofilm grown in the CDC biofilm reactor following the procedures outlined in Practice E3161. Biofilm grown in the CDC reactor is representative of biofilm that forms under high fluid shear on surfaces conducive to biofilm formation.  
5.1.1 Vegetative biofilm bacteria are phenotypically different from suspended planktonic cells of the same genotype. Biofilm growth reactors are engineered to produce biofilm with specific characteristics (2). Altering either the engineered system or operating conditions will modify those characteristics as well as the physicochemical environment. The goal in biofilm research and testing is to choose the growth reactor and operating conditions that generate the most relevant biofilm for the particular study.  
5.2 The test method was designed to determine the log10 reduction in bacteria after exposure to a disinfectant in a closed system.  
5.3 The test method uses 50 mL conical tubes. The conical geometry allows for disinfectant exposure to biofilm on all surfaces of the coupon. For foaming disinfectants or for disinfectants requiring a larger volume of neutralizer, 250 mL conical tubes are used which preserve the required geometry and allow for greater neutralization capacity.  
5.4 Each test includes three untreated control coupons (exposed to buffered dilution water) and five treated coupons (per disinfectant/concentration/contact time combination).
SCOPE
1.1 This test method specifies the operational parameters required to perform a quantitative liquid disinfectant efficacy test against bacterial biofilm.  
1.2 The test method was optimized and validated for a Pseudomonas aeruginosa or Staphylococcus aureus biofilm grown in the CDC Biofilm Reactor (E3161). The method is suitable for evaluating additional bacteria grown using the procedures outlined in methods with comparable coupon dimensions such as Practice E3161, Test Method E2562, or Test Method E2196.  
1.3 Disinfectant preparation and contact time are used in the assessment according to the manufacturer’s instructions for use.  
1.4 The test method uses a closed system to treat biofilm. A coupon is placed in a single tube for the treatment, neutralization, and harvesting steps to prevent the loss of cells.  
1.5 This test method describes a harvesting and analysis procedure which includes vortexing and sonicating treated and untreated control biofilm, and recovery of culturable cells using filtration to lower the limit of detection. Biofilm population density is recorded as log10 colony-forming units per coupon. Efficacy is reported as a log10 reduction of culturable cells.  
1.6 Basic microbiology training is required to perform this assay.  
1.7 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.8 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.  
1.9 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

• Status: Published
• Publication Date: 31-Oct-2021
• ICS: 71.100.35 - Chemicals for industrial and domestic disinfection purposes
• Technical Committee: E35 - Pesticides, Antimicrobials, and Alternative Control Agents
• Drafting Committee: E35.15 - Antimicrobial Agents

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:E2871 −21
Standard Test Method for
Determining Disinfectant Efficacy Against Biofilm Grown in
1
the CDC Biofilm Reactor Using the Single Tube Method
This standard is issued under the fixed designation E2871; 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* 1.9 This international standard was developed in accor-
dance with internationally recognized principles on standard-
1.1 This test method specifies the operational parameters
ization established in the Decision on Principles for the
required to perform a quantitative liquid disinfectant efficacy
Development of International Standards, Guides and Recom-
test against bacterial biofilm.
mendations issued by the World Trade Organization Technical
1.2 The test method was optimized and validated for a
Barriers to Trade (TBT) Committee.
Pseudomonas aeruginosa or Staphylococcus aureus biofilm
grown in the CDC Biofilm Reactor (E3161). The method is 2. Referenced Documents
suitable for evaluating additional bacteria grown using the 2
2.1 ASTM Standards:
procedures outlined in methods with comparable coupon
E2196Test Method for Quantification of Pseudomonas
dimensions such as Practice E3161, Test Method E2562,or
aeruginosa Biofilm Grown with Medium Shear and
Test Method E2196.
Continuous Flow Using Rotating Disk Reactor
1.3 Disinfectantpreparationandcontacttimeareusedinthe
E2562Test Method for Quantification of Pseudomonas
assessment according to the manufacturer’s instructions for aeruginosaBiofilmGrownwithHighShearandContinu-
use.
ous Flow using CDC Biofilm Reactor
E2756Terminology Relating toAntimicrobial andAntiviral
1.4 The test method uses a closed system to treat biofilm.A
Agents
coupon is placed in a single tube for the treatment,
E3161PracticeforPreparinga Pseudomonas aeruginosaor
neutralization, and harvesting steps to prevent the loss of cells.
Staphylococcus aureus Biofilm using the CDC Biofilm
1.5 This test method describes a harvesting and analysis
Reactor
procedure which includes vortexing and sonicating treated and
2.2 Other Standards:
untreated control biofilm, and recovery of culturable cells
Method 9050 C.1.a Buffered Dilution Water Preparation
using filtration to lower the limit of detection. Biofilm popu-
3
according to Eaton et al (1)
lation density is recorded as log colony-forming units per
10
coupon. Efficacy is reported as a log reduction of culturable
3. Terminology
10
cells.
3.1 For definitions of terms used in this test method refer to
1.6 Basic microbiology training is required to perform this
Terminology E2756.
assay.
3.2 Definitions:
1.7 The values stated in SI units are to be regarded as
3.2.1 biofilm, n—microorganisms living in a self-organized
standard. No other units of measurement are included in this
community attached to surfaces, interfaces, or each other,
standard.
embedded in a matrix of extracellular polymeric substances of
microbial origin, while exhibiting altered phenotypes with
1.8 This standard does not purport to address all of the
respect to growth rate and gene transcription.
safety concerns, if any, associated with its use. It is the
3.2.1.1 Discussion—Biofilm may be comprised of bacteria,
responsibility of the user of this standard to establish appro-
fungi, algae, protozoa, viruses, or infinite combinations of
priate safety, health, and environmental practices and deter-
these microorganisms. The qualitative characteristics of a
mine the applicability of regulatory limitations prior to use.
1 2
This test method is under the jurisdiction of ASTM Committee E35 on For referenced ASTM standards, visit the ASTM website, www.astm.org, or
Pesticides, Antimicrobials, and Alternative Control Agents and is the direct contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM
responsibility of Subcommittee E35.15 on Antimicrobial Agents. Standards volume information, refer to the standard’s Document Summary page on
Current edition approved Nov. 1, 2021. Published January 2022. Originally the ASTM website.
3
approved in 2012. Last previous edition approved in 2019 as E2871–19. DOI: The boldface numbers in parentheses refer to a list of references at the end of
10.1520/E2871–21. this standard.
*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

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E2871−21
biofilm inc
...

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: E2871 − 19 E2871 − 21
Standard Test Method for
Determining Disinfectant Efficacy Against Biofilm Grown in
1
the CDC Biofilm Reactor Using the Single Tube Method
This standard is issued under the fixed designation E2871; 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 Scope*
1.1 This test method specifies the operational parameters required to perform a quantitative liquid disinfectant efficacy test against
bacterial biofilm.
1.2 The test method was optimized and validated for a Pseudomonas aeruginosa or Staphylococcus aureus biofilm grown in the
CDC Biofilm Reactor (E3161). The method is suitable for evaluating additional bacteria grown using the procedures outlined in
methods with comparable coupon dimensions such as Practice E3161, Test Method E2562, or Test Method E2196.
1.3 Disinfectant preparation and contact time are used in the assessment according to the manufacturer’s instructions for use.
1.4 The test method uses a closed system to treat biofilm. A coupon is placed in a single tube for the treatment, neutralization,
and harvesting steps to prevent the loss of cells.
1.5 This test method describes a harvesting and analysis procedure which includes vortexing and sonicating treated and untreated
control biofilm, and recovery of culturable cells using filtration to lower the limit of detection. Biofilm population density is
recorded as log colony-forming units per coupon. Efficacy is reported as a log reduction of culturable cells.
10 10
1.6 Basic microbiology training is required to perform this assay.
1.7 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.8 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.
1.9 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.
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 Jan. 1, 2019Nov. 1, 2021. Published February 2019January 2022. Originally approved in 2012. Last previous edition approved in 20132019 as
E2871–13.–19. DOI: 10.1520/E2871–19.10.1520/E2871–21.
*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 ----------------------
E2871 − 21
2. Referenced Documents
2
2.1 ASTM Standards:
E1054 Practices for Evaluation of Inactivators of Antimicrobial Agents
E2196 Test Method for Quantification of Pseudomonas aeruginosa Biofilm Grown with Medium Shear and Continuous Flow
Using Rotating Disk Reactor
E2562 Test Method for Quantification of Pseudomonas aeruginosa Biofilm Grown with High Shear and Continuous Flow using
CDC Biofilm Reactor
E2756 Terminology Relating to Antimicrobial and Antiviral Agents
E3161 Practice for Preparing a Pseudomonas aeruginosa or Staphylococcus aureus Biofilm using the CDC Biofilm Reactor
2.2 Other Standards:
3
Method 9050 C.1.a Buffered Dilution Water Preparation according to Eaton et al (1)
3. Terminology
3.1 For definitions of terms used in this test method refer to Terminology E2756.
3.2 Definitions:
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—
Biofilm 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 dive
...