Enumeration procedure for monitoring test microbe populations on inoculated carriers in AOAC use-dilution methods.
The AOAC Use-Dilution methods do not provide procedures to enumerate the test microbe on stainless steel carriers (penicylinders) or guidance on the expected target populations of the test microbe (i.e., a performance standard). This report describes the procedures used by the U.S. Environmental Protection Agency to enumerate the test microbe (carrier counts) associated with conducting the Use-Dilution method with Staphylococcus aureus (Method 955.15) and Pseudomonas aeruginosa (Method 964.02) and the examination of historical data. The carrier count procedure involves the random selection of carriers, shearing bacterial cells from the carrier surface through sonication, and plating of serially diluted inoculum on trypticase soy agar. For each Use-Dilution test conducted, the official AOAC method was strictly followed for carrier preparation, culture initiation, test culture preparation, and carrier inoculation steps. Carrier count data from 78 Use-Dilution tests conducted over a 6-year period were compiled and analyzed. A mean carrier count of 6.6 logs (approximately 4.0 x 10(6) colony-forming units/carrier) was calculated for both S. aureus and P. aeruginosa. Of the mean values, 95% fell within +/- 2 repeatability standard deviations. The enumeration procedure and target carrier counts are desirable for standardizing the Use-Dilution methods, increasing their reproducibility, and ensuring the quality of the data.
Tomasino
Stephen F
SF
U.S. Environmental Protection Agency, Office of Pesticide Programs, Microbiology Laboratory, Environmental Science Center, Fort Meade, MD 20755-5350, USA. tomasino.stephen@epa.gov
Fiumara
Rebecca M
RM
Cottrill
Michele P
MP
D015169:Colony Count, Microbial / Q000379:methods*; D003470:Culture Media; D007201:Indicator Dilution Techniques; D007202:Indicators and Reagents; D011550:Pseudomonas aeruginosa; D011786:Quality Control; D013211:Staphylococcus aureus; D014481:United States; D014484:United States Environmental Protection Agency
D003470:Culture Media; D007202:Indicators and Reagents