Background: The American Heart Association first developed recommendations on antibiotic prophylaxis against infective endocarditis during dental procedures more than 50 years ago. These recommendations were partly based on the fact that bacteremia occurs with dental procedures. Previous studies in the 1970s and earlier demonstrated that patients become bacteremic after tooth brushing. Improved culture techniques suggest that these rates could be higher now. The objective of this study was to determine the current incidence of bacteremia after routine tooth brushing. Methods: Thirty military beneficiaries were enrolled in a prospective, institutional review board-approved study after providing informed consent. The incidence of bacteremia after routine tooth brushing for 1 minute using a standardized soft-bristle toothbrush was prospectively measured in 30 healthy adults at three different time points (at baseline and 30 seconds and 20 minutes after brushing). Periodontal Screening and Recording (PSR) scores were recorded for each patient to assess periodontal disease. Results: Three of 180 blood cultures were positive for Propionibacterium acnes (a known contaminant). The remaining blood cultures were all negative. The average PSR score was 9.8 (standard deviation 3.17) for 17 of 30 subjects. Conclusions: The rate of true bacteremia in this study was zero, which is much lower than previous studies. Bacteremia after tooth brushing in a healthy population is a rare occurrence. Data from previous studies may no longer apply to the current population. Results similar to the ones found in this study during other dental procedures could be an impetus to reevaluate infective endocarditis prophylaxis guidelines.