Quantitative evaluation of bacteria washed from stainless steel penicylinders during AOAC use-dilution method

Stainless steel penicylinders inoculated separately with test bacteria (Salmonella choleraesuis, Pseudomonas aeruginosa, or Staphylococcus aureus) are used in the AOAC use-dilution method (UDM) for disinfectant efficacy testing. Numbers of bacteria remaining on penicylinders were quantitatively assessed to determine if cells are washed from the penicylinders after a 10 min exposure to phosphate buffer dilution water (PBDW). Inoculated penicylinders were also examined by scanning electron microscopy (SEM) to determine the presence of cells remaining attached to the penicylinders after a 10 min exposure to a quaternary ammonium disinfectant and separately to PBDW. The percentage of cells washed from inoculated penicylinders exposed to PBDW was 89.9 for Salmonella choleraesuis, 48.8 for Pseudomonas aeruginosa, and 38.8 for Staphylococcus aureus. Qualitative examination of penicylinders by scanning electron microscopy confirmed the attachment of S. aureus and P. aeruginosa cells to penicylinders exposed separately to PBDW and a quaternary ammonium disinfectant. Few S. choleraesuis cells were observed on penicylinders exposed to PBDW and no cells were observed after disinfectant exposure. The variability of the numbers of viable cells entering the recovery media among the 3 UDM test bacteria due to cell detachment could be a significant factor in the recognized variability of the use-dilution method.