Browsing by Author "Cottrill, M. P."

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Determining the efficacy of liquid sporicides against spores of Bacillus subtilis on a hard nonporous surface using the quantitative three step method: Collaborative study
(2008-07) Tomasino, S. F.; Pines, R. M.; Cottrill, M. P.; Hamilton, Martin A.
A collaborative study was conducted to validate the quantitative Three Step Method (TSM), a method designed to measure the performance of liquid sporicides on a hard nonporous surface. Ten laboratories agreed to participate in the collaborative study; data from 8 of 10 participating laboratories were used in the final statistical analysis. The TSM uses 5 x 5 x 1 mm glass coupons (carriers) upon which spores have been inoculated and which are introduced into liquid sporicidal agent contained in a microcentrifuge tube. Following exposure to a test chemical and a neutralization agent, spores are removed from carriers in 3 fractions: passive removal (Fraction A), sonication (Fraction B), and gentle agitation (Fraction C). Liquid from each fraction is serially diluted and plated on a recovery medium for spore enumeration. Control counts are compared to the treated counts, and the level of efficacy is determined by calculating the log10 reduction (LR) of spores. The main statistical goals were to evaluate the repeatability and reproducibility of the LR values, to estimate the components of variance for LR, and to assess method responsiveness. AOAC Method 966.04â€“Method II was used as a reference method. The scope of the validation was limited to testing liquid formulations against spores of Bacillus subtilis, a surrogate for virulent strains of B. anthracis, on a hard nonporous surface (glass). The test chemicals used in the study were sodium hypochlorite, a combination of peracetic acid and hydrogen peroxide, and glutaraldehyde. Each test chemical was evaluated at 3 levels of presumed efficacy: high, medium, and low. Three replications were required. The TSM was validated as it successfully met the statistical parameters for quantitative test methods. Satisfactory validation parameters, such as the repeatability standard deviation (Sr) and reproducibility standard deviation (SR), were obtained for control carrier counts and LR values. Both the TSM and the reference method were responsive to the efficacy levels of the test chemicals. For the 72 total TSM tests conducted, the mean (Â± standard error of the mean) log density of spores per control carrier was 6.86 (Â± 0.08); the Sr and SR were low at 0.15 and 0.27, respectively. Across the range of test chemicals, the Sr and SR estimates associated with LR were also acceptably low. The Sr ranged from 0.17 to 0.72 and the SR ranged from 0.34 to 1.43. Overall, the Sr and SR estimates associated with the efficacy data were within the ranges published for other quantitative methods and meet the performance characteristics necessary for validation. Collaborators: Alvey K; Buen M; Chan-Myers H; Chang G; Dellâ€™Aringa B; Gonzales E; Hitchins V; Hollingsworth A; Jeske A; Kingma D; Kitchen nee Dormstetter K; Klein D; Lappalainen S; Lawrence J; Lehman L; Malulla K; Michler T; Paulson D; Regan P; Rodriguez A; Rottjakob D; Sathe M; Steinagel S; Suchmann D; Tester J; To T; Wieland D; Zhang Q
Procedural revision to the AOAC germicidal spray products as disinfectants test method: Establishment of minimum and maximum log density values for test microbes on inoculated carriers
(2013-06) Pines, R. M.; Tomasino, S. F.; Cottrill, M. P.; Hamilton, G. C.; Parker, Albert E.
The AOAC Germicidal Spray Products as Disinfectants test method (AOAC Official Method 961.02) is used to measure the efficacy of spray products on hard inanimate surfaces; however, the method does not provide procedures to determine the population of the test microbe on inoculated glass slide carriers (e.g., carrier counts reported as CFU/carrier). 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 issue and, based on carrier count data collected by four laboratories from 2000 to 2010, proposes a specific range for the mean log density per carrier as a requirement. Laboratory-based carrier count data were collected concurrently with 116 Method 961.02 efficacy tests conducted on spray products bearing claims against Pseudomonas aeruginosa and Staphylococcus aureus. For many of the tests a soil load (SL) was added to the inoculum (as specified on the product label claim). Six carriers were assayed per test for a total of 696 carriers. All but two of the 116 mean log densities were at least 5.0 (a geometric mean of 1.0 Ã— 105 CFU/carrier). Across the four combinations of microbes and SL treatments, the mean TestLD (mean log density across all enumerated carriers in a test) ranged from approximately 6.0 (a geometric mean of 0.9 Ã— 106 CFU/carrier) to 6.3 (a geometric mean of 2.0 Ã— 106 CFU/carrier). Across all microbes and SL treatments, the mean log density (Â±SEM) was 6.2 (Â±0.07) per carrier (a geometric mean of 1.5 Ã— 106 CFU/carrier). The mean log density for six carriers per test showed good repeatability (0.32) and reproducibility (0.34). The proposed requirement for S. aureus tests and P. aeruginosa tests is a mean log density (across six carriers) between 5.0 and 6.5. A separate 2009 study at three laboratories was conducted to evaluate the persistence of P. aeruginosa, S. aureus, and Salmonella enterica on glass carriers. Based on the persistence data, a 2 h use period is proposed for using the inoculated carriers post drying. The persistence data set was also used to assess the carrier counts for S. enterica. The carrier counts were approximately one log lower for S. enterica compared to S. aureus and P. aeruginosa; a range of 4.0 to 5.5 logs is proposed as a requirement for S. enterica tests.