ASTM_E_2799_-_12.pdf

Designation:E279912Standard Test Method forTesting Disinfectant Efficacy against Pseudomonasaeruginosa Biofilm using the MBEC Assay1This standard is issued under the fixed designation E2799;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 grow and treat a Pseudomonas aeruginosa biofilmin a high throughput screening assay known as the MBEC(trademarked)2(Minimum Biofilm Eradication Concentration)Physiology and Genetics Assay.The assay device consists of aplastic lid with ninety-six(96)pegs and a correspondingreceiver plate with ninety-six(96)individual wells that have amaximum 200-L working volume.Biofilm is established onthe pegs under batch conditions(that is,no flow of nutrientsinto or out of an individual well)with gentle mixing.Theestablished biofilm is transferred to a new receiver plate fordisinfectant efficacy testing.3,4The reactor design allows forthe simultaneous testing of multiple disinfectants or onedisinfectant with multiple concentrations,and replicatesamples,making the assay an efficient screening tool.1.2 This test method defines the specific operational param-eters necessary for growing a Pseudomonas aeruginosabiofilm,although the device is versatile and has been used forgrowing,evaluating and/or studying biofilms of differentspecies as seen in Refs(1-4).51.3 Validation of disinfectant neutralization is included aspart of the assay.1.4 This test method describes how to sample the biofilmand quantify viable cells.Biofilm population density is re-corded as log10colony forming units per surface area.Efficacyis reported as the log10reduction of viable cells.1.5 Basic microbiology training is required to perform thisassay.1.6 The values stated in SI units are to be regarded asstandard.No other units of measurement are included in thisstandard.1.7 ASTM International takes no position respecting thevalidity of any patent rights asserted in connection with anyitem mentioned in this standard.Users of this standard areexpressly advised that determination of the validity of any suchpatent rights,and the risk of infringement of such rights,areentirely their own responsibility.1.8 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:6E1054 Test Methods for Evaluation of Inactivators of Anti-microbial Agents2.2 Other Standards:Method 9050 C.1.a Buffered Dilution Water Preparationaccording to Eaton et al(5)3.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 DiscussionBiofilms may be comprised of bacteria,fungi,algae,protozoa,viruses,or infinite combinations of1This 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 April 1,2012.Published June 2012.Originallyapproved in 2011.Last previous edition approved in 2011 as E2799 11.DOI:10.1520/E279912.2The MBEC trademark is held by Innovotech,Inc.,Edmonton,Alberta,Canada.3The sole source of supply of the apparatus known to the committee at this timeis Innovotech Inc.,Edmonton,Alberta,Canada.If you are aware of alternativesuppliers,please provide this information to ASTM International Headquarters.Your comments will receive careful consideration at a meeting of the responsibletechnical committee,1which you may attend.4The MBECAssay is covered by a patent.Interested parties are invited to submitinformation regarding the identification of an alternative(s)to this patented item tothe ASTM International Headquarters.Your comments will receive careful consid-eration at a meeting of the responsible technical committee,1which you may attend.5The boldface numbers in parentheses refer to a list of references at the end ofthis standard.6For 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.Copyright ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959.United States1 these microorganisms.The qualitative characteristics of abiofilm including,but not limited to,populatio