ASTM_E_2871_-_13.pdf

Designation:E287113Standard Test Method forEvaluating Disinfectant Efficacy Against Pseudomonasaeruginosa Biofilm Grown in CDC Biofilm Reactor UsingSingle Tube Method1This standard is issued under the fixed designation E2871;the number immediately following the designation indicates the year oforiginal adoption or,in the case of revision,the year of last revision.A number in parentheses indicates the year of last reapproval.Asuperscript epsilon()indicates an editorial change since the last revision or reapproval.1.Scope1.1 This test method specifies the operational parametersrequired to perform a quantitative liquid disinfectant efficacytest against biofilm bacteria.1.2 The test method was developed using a Pseudomonasaeruginosa biofilm grown in the CDC Biofilm Reactor(TestMethod E2562),modified to include borosilicate glass couponsas a hard nonporous surface and P.aeruginosa ATCC 15442.1.3 Disinfectant preparation and contact time are used in theassessment according to the manufacturers instructions foruse.1.4 The test method uses a closed system to treat biofilm.Acoupon is placed in a single tube for the treatment,neutralization,and sampling steps to prevent the loss of cells.1.5 Verification of disinfectant neutralization is determinedprior to conducting the test method.1.6 This test method describes how to sample and analyzetreated and untreated control biofilms for viable cells.Biofilmpopulation density is recorded as log10colony-forming unitsper coupon.Efficacy is reported as a log10reduction of viablecells.1.7 Basic microbiology training is required to perform thisassay.1.8 The values stated in SI units are to be regarded asstandard.No other units of measurement are included in thisstandard.1.9 This standard does not purport to address all of thesafety concerns,if any,associated with its use.It is theresponsibility of the user of this standard to establish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use.2.Referenced Documents2.1 ASTM Standards:2E1054 Test Methods for Evaluation of Inactivators of Anti-microbial AgentsE2562 Test Method for Quantification of Pseudomonasaeruginosa Biofilm Grown with High Shear and Continu-ous Flow using CDC Biofilm Reactor2.2 Other Standards:Method 9050 C.1.a Buffered Dilution Water Preparationaccording to Eaton et al(1)33.Terminology3.1 Definitions:3.1.1 biofilm,nmicroorganisms living in a self-organizedcommunity attached to surfaces,interfaces,or each other,embedded in a matrix of extracellular polymeric substances ofmicrobial origin,while exhibiting altered phenotypes withrespect to growth rate and gene transcription.3.1.1.1 DiscussionBiofilm may be comprised of bacteria,fungi,algae,protozoa,viruses,or infinite combinations ofthese microorganisms.The qualitative characteristics of abiofilm including,but not limited to,population density,taxonomic diversity,thickness,chemical gradients,chemicalcomposition,consistency,and other materials in the matrix thatare not produced by the biofilm microorganisms,are controlledby the physicochemical environment in which it exists.3.1.2 contact time,npredetermined time that the biofilm isexposed to the activity of a disinfectant.3.1.3 coupon,nbiofilm growth surface.3.1.4 disinfectant,na chemical that destroys vegetativeforms of microorganisms,but does not ordinarily kill bacterialspores.3.2 Acronyms:1This test method is under the jurisdiction of ASTM Committee E35 onPesticides,Antimicrobials,and Alternative Control Agents and is the directresponsibility of Subcommittee E35.15 on Antimicrobial Agents.Current edition approved Oct.1,2013.Published November 2013.Originallyapproved in 2012.Last previous edition approved in 2012 as E287112.DOI:10.1520/E287113.2For referenced ASTM standards,visit the ASTM website,www.astm.org,orcontact ASTM Customer Service at serviceastm.org.For Annual Book of ASTMStandards volume information,refer to the standards Document Summary page onthe ASTM website.3The boldface numbers in parentheses refer to a list of references at the end ofthis standard.Copyright ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959.United States1 3.2.1 ATCCAmerican Type Culture Collection.3.2.2 CDCCenters for Disease Control and Prevention.3.2.3 CFUcolony-forming unit.4.Summary of Test Method4.1 This test method describes the use of the single tubemethod to evaluate the efficacy of a liquid disinfectant againsta Pseudomonas aeruginosa biofilm on a hard nonporoussurface grown in the CDC Biofilm Reactor.The test methodconsists of adding a disinfectant(treated)or a control buffer(untreated)to individual coupons held in 50-mL conical tubes.Three coupons are treated with disinfectant and three couponsreceive buffered dilution water.Neutralizer is added to thetubes after the appropriate contact time.A combination ofvortexing and sonication are used to remove the biofilm fromthe coupon and disaggregate the clumps.The cell suspension isser