title: Ruggedness and reproducibility of the MBEC biofilm disinfectant efficacy test.
abstract: The MBEC™ Physiology & Genetics Assay recently became the first approved ASTM standardized biofilm disinfectant efficacy test method. This report summarizes the results of the standardization process using Pseudomonas aeruginosa biofilms. Initial ruggedness testing of the MBEC method suggests that the assay is rugged (i.e., insensitive) to small changes to the protocol with respect to 4 factors: incubation time of the bacteria (when varied from 16 to 18h), treatment temperature (20-24°C), sonication duration (25-35min), and sonication power (130-480W). In order to assess the repeatability of MBEC results across multiple tests in the same laboratory and the reproducibility across multiple labs, an 8-lab study was conducted in which 8 concentrations of each of 3 disinfectants (a non-chlorine oxidizer, a phenolic, and a quaternary ammonium compound) were applied to biofilms using the MBEC method. The repeatability and reproducibility of the untreated control biofilms were acceptable, as indicated by small repeatability and reproducibility standard deviations (SD) (0.33 and 0.67 log10(CFU/mm(2)), respectively). The repeatability SDs of the biofilm log reductions after application of the 24 concentration and disinfectant combinations ranged from 0.22 to 1.61, and the reproducibility SDs ranged from 0.27 to 1.70. In addition, for each of the 3 disinfectant types considered, the assay was statistically significantly responsive to the increasing treatment concentrations.
authors.affiliation: Center for Biofilm Engineering, Montana State University, Bozeman, MT 59715, USA; Department of Mathematical Sciences, Montana State University, Bozeman, MT 59715, USA. Electronic address: parker@math.montana.edu.
authors.forename: A E
authors.initials: AE
authors.lastname: Parker
authors.affiliation: Center for Biofilm Engineering, Montana State University, Bozeman, MT 59715, USA.
authors.forename: D K
authors.initials: DK
authors.lastname: Walker
authors.affiliation: Center for Biofilm Engineering, Montana State University, Bozeman, MT 59715, USA.
authors.forename: D M
authors.initials: DM
authors.lastname: Goeres
authors.affiliation: Innovotech Inc., Edmonton AB T6N 1H1, Canada.
authors.forename: N
authors.initials: N
authors.lastname: Allan
authors.affiliation: Innovotech Inc., Edmonton AB T6N 1H1, Canada.
authors.forename: M E
authors.initials: ME
authors.lastname: Olson
authors.affiliation: Innovotech Inc., Edmonton AB T6N 1H1, Canada.
authors.forename: A
authors.initials: A
authors.lastname: Omar
mesh_terms: D018441:Biofilms / Q000187:drug effects*; D004202:Disinfectants / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D008826:Microbial Sensitivity Tests / Q000379:methods; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000502:physiology*; D015203:Reproducibility of Results; D013010:Sonication; D013696:Temperature; D013997:Time Factors
chemical_list: D004202:Disinfectants
keywords: Biofilm; Reproducibility; Ruggedness