title: Improving the AOAC use-dilution method by establishing a minimum log density value for test microbes on inoculated carriers.
abstract: The AOAC Use-Dilution methods, 955.14 (Salmonella enterica), 955.15 (Staphylococcus aureus), and 964.02 (Pseudomonas aeruginosa), are used to measure the efficacy of disinfectants on hard inanimate surfaces. The methods do not provide procedures to assess log density of the test microbe on inoculated penicylinders (carrier counts). Without a method to measure and monitor carrier counts, the associated efficacy data may not be reliable and repeatable. This report provides a standardized procedure to address this method deficiency. Based on carrier count data collected by four laboratories over an 8 year period, a minimum log density value is proposed to qualify the test results. Carrier count data were collected concurrently with 242 Use-Dilution tests. The tests were conducted on products bearing claims against P. aeruginosa and S. aureus with and without an organic soil load (OSL) added to the inoculum (as specified on the product label claim). Six carriers were assayed per test for a total of 1452 carriers. All 242 mean log densities were at least 6.0 (geometric mean of 1.0 x 10(6) CFU/carrier). The mean log densities did not exceed 7.5 (geometric mean of 3.2 x 10(7) CFU/carrier). For all microbes and OSL treatments, the mean log density (+/- SEM) was 6.7 (+/- 0.07) per carrier (a geometric mean of 5.39 x 10(6) CFU/carrier). The mean log density for six carriers per test showed good repeatability (0.29) and reproducibility (0.32). A minimum mean log density of 6.0 is proposed as a validity requirement for S. aureus and P. aeruginosa. The minimum level provides for the potential inherent variability that may be experienced by a wide range of laboratories and the slight effect due to the addition of an OSL. A follow-up report is planned to present data to support the carrier count procedure and carrier counts for S. enterica.
authors.affiliation: U.S. Environmental Protection Agency, Office of Pesticide Programs, Microbiology Laboratory, Environmental Science Center, Fort Meade, MD 20755-5350, USA. tomasino.stephen@epa.gov
authors.forename: Stephen F
authors.initials: SF
authors.lastname: Tomasino
authors.forename: Rebecca M
authors.initials: RM
authors.lastname: Pines
authors.forename: Martin A
authors.initials: MA
authors.lastname: Hamilton
mesh_terms: D002623:Chemistry Techniques, Analytical / Q000379:methods*; D015169:Colony Count, Microbial / Q000379:methods*; D003470:Culture Media; D007201:Indicator Dilution Techniques; D007202:Indicators and Reagents; D011550:Pseudomonas aeruginosa / Q000378:metabolism*; D011786:Quality Control; D019779:Salmonella enterica / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000378:metabolism*; D014481:United States; D014484:United States Environmental Protection Agency
chemical_list: D003470:Culture Media; D007202:Indicators and Reagents