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Finally, an increasing inoculum size in log-phase cultures is approaching the stationary growth phase and hence these organisms may be killed more slowly. Thus, both inoculum size and growth phase may independently increase the rate of killing. The inoculum size is considered the single most important variable in susceptibility testing. The inoculum size is critical; variations in density of inoculum may alter then endpoints by one or more dilutions, and therefore the preparation of a standardized inoculum is critical. NCCLS has recommended a final inoculum size of 5 x 105 CFU/mL for susceptibility testing of aerobic bacteria in broth.2

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