Identifying the factors shaping dialects can reveal selective pressures and cultural influences on acoustic signals. Translocations of species have been considered the “gold standard” for identifying population differences in vocalizations, including dialects. The North American elk (Cervus canadensis) is a species that has experienced numerous translocations, but no published studies exist on dialects in elk or any other ungulate species. Adult male North American elk utter an iconic vocalization during the breeding season, which is termed the bugle call due to the power, duration, and pitch of the sound. We investigated if dialect differences existed between three populations: a source population in Wyoming (N = 10 bulls, 132 calls) and translocated populations in Colorado (N = 13 bulls, 92 calls) and Pennsylvania (N = 20 bulls, 160 calls). Dialect differences existed between the populations in 9 of 10 measured variables, in both the frequency domain and time domain. Habitat factors (acoustic adaptation hypothesis) affecting sound transmission were inconsistently related to dialects in the populations. Genetic factors may be related to dialects, as the historically bottlenecked Pennsylvania population, with low heterozygosity and low allelic richness, exhibited the least variability in acoustic measures. Cultural transmission and vocal learning may also contribute to dialect development in this highly vocal, social ungulate species.