The aim of this study was to establish the incidence of fecal carriage of Listeria monocytogenes in healthy adults. A total of 868 stool specimens from three healthy volunteers (1 male and 2 females; ages 44, 39, and 60 years) were collected between 1 October 2000 and 30 September 2001. Culture was performed using Fraser broth and Palcam selective agar plates. Polymerase chain reaction (PCR) was performed using Probelia Listeria monocytogenes (BioRad, France). Overall, Listeria monocytogenes was detected in 31 of the 868 (3.57%) stool specimens using PCR. Sixteen of the 31 positive results were single events, i.e., samples collected from the same patient the day before and the day after the positive result were both negative. Positive results on two consecutive days were found four times, on three consecutive days one time, and on four consecutive days one time. Listeria monocytogenes was cultured from 10 of 868 (1.15%) stool specimens. These culture-positive samples, all positive by PCR as well, accounted for five independent episodes. Using automated ribotyping on up to 40 single colonies per stool specimen, 9 of 10 culture-positive samples yielded more than one strain. There was no obvious seasonal clustering of positive results. None of the documented episodes of Listeria monocytogenes carriage, all of which involved serotypes 1/2a and 1/2b, coincided with overt illness. The results of PCR indicate an incidence of five to nine exposures to Listeria monocytogenes per person per year. On average, the incidence of culture-confirmed fecal carriage in healthy adults is two episodes of Listeria monocytogenes carriage per person per year. Fecal shedding was of short duration (maximum 4 days), which argues against the appropriateness of routine stool screening in dairy workers as a tool for prevention of listeriosis.