A multiyear survey of 31 ready-to-eat (RTE) food processing plants in the United States was conducted to determine the incidence of Listeria spp. in various RTE production environments. Samples were collected from 22 RTE plants regulated by the U.S. Department of Agriculture’s Food Safety and Inspection Service (FSIS) and from 9 RTE food plants regulated by the U.S. Department of Health and Human Services’ Food and Drug Administration (FDA). Only nonfood contact surfaces in the RTE manufacturing areas with exposed RTE product were sampled. Each sample was individually analyzed for the presence of Listeria spp. by using a PCR-based rapid assay. In total, 4,829 samples were collected from various locations, including freezers, equipment framework, floors, walls, wall-floor junctures, drains, floor mats, doors, and cleaning tools. Nine (29%) of the facilities had zero samples positive for Listeria spp. in the production environment, whereas 22 (71%) had one or more samples positive for Listeria spp. The total incidence of Listeria spp. in all RTE food plants was 4.5%. The positive rate in plants regulated by the FSIS ranged from 0 to 9.7%, whereas the positive rate in plants regulated by the FDA ranged from 1.2 to 36%.