Two juvenile scimitar-horned oryx (Oryx dammah) at the Wild Animal Park Planckendael died from acute septicemia caused by Listeria monocytogenes serovar 4b. Subsequently, Listeria spp. were isolated from the feces, food, and environment of seven antelope species and examined using a two-stage enrichment procedure in Fraser Broth, followed by isolation on PALCAM agar. A total of 40/170 samples (23.5%) was positive for Listeria spp. No organisms were cultured in 83/170 samples (48.8%), and 47 samples (27.6%) were overgrown with Bacillus spp. Nonpathogenic Listeria spp. were isolated from 16/70 fecal samples, 22/40 soil samples, and 2/60 feed samples. Listeria monocytogenes serovar 1/2b was isolated from two soil samples collected in the enclosure of the scimitar-horned oryx.