This study compared the effectiveness of hand and ultrasonic instrumentation for removing a standardized inoculum of pigmented Serratia marcescens from the root canal system of premolars in dogs. Forty-four premolars from nine beagle dogs were divided into two experimental groups of 20 and 24 teeth, respectively. The experimental teeth were inoculated with approximately 10 colony-forming units of S. marcescens. After the bacterial were allowed to colonize for 1 wk, the experimental teeth were instrumented with either hand instruments or the Cavi-Endo device. The teeth were extracted, crushed, and assayed for recoverable colony-forming units of S. marcescens. Statistical comparisons of the ratio of inoculated to recovered colony-forming untis were made. The results indicated that the difference between the positive controls and the experimental groups was significant. There was no significant difference between the two instrumentation groups.