Antibiotics used to reduce nasal colonization by Staphylococcus aureus in patients before admission are inappropriate for carriage reduction on a regular basis within a hospital community. Effective nonantibiotic alternatives for daily use in the nares will allow reduction of this bacterial source to be addressed. Our study tested the effectiveness of a nonantibiotic, alcohol-based antiseptic in reducing nasal bacterial carriage in health care professionals (HCPs) at an urban hospital center. HCPs testing positive for vestibular S aureus colonization were treated 3 times during the day with topical antiseptic or control preparations. Nasal S aureus and total bacterial colonization levels were determined before and at the end of a 10-hour workday. Seventy-eight of 387 HCPs screened (20.2%) tested positive for S aureus infection. Of 39 subjects who tested positive for S aureus infection who completed the study, 20 received antiseptic and 19 received placebo treatment. Antiseptic treatment reduced S aureus colony forming units from baseline by 99% (median) and 82% (mean) (P < .001). Total bacterial colony forming units were reduced by 91% (median) and 71% (mean) (P < .001). Nasal application of a nonantibiotic, alcohol-based antiseptic was effective in reducing S aureus and total bacterial carriage, suggesting the usefulness of this approach as a safe, effective, and convenient alternative to antibiotic treatment.