A few species of the weakly electric snoutfish, the African freshwater family Mormyridae, have been reported to vocalise. However, allopatric populations of a single species were never compared. Members of three allopatric Marcusenius macrolepidotus populations, originating from the Upper Zambezi River in Namibia, the Buzi River (Mozambique), and the Incomati River system in South Africa, vocalised with pulsatile growl- and tonal hoot sounds in dyadic confrontation experiments. A high rate of growling accompanied territorial and agonistic interactions and also non-threatening interactions between males and females, which in one pair appeared to be courtship. Growl sound characteristics of M. macrolepidotus from the Incomati system differed from those of the Upper Zambezi in a significantly higher frequency of the first harmonic (mean, 355 Hz vs 266 Hz). The two vocalising males from the Buzi River generated growls about twice as long as the other fish. Furthermore, the growl pulse period was about 4 ms in M. macrolepidotus from the Upper Zambezi River and from the Incomati system, but 6 ms in M. macrolepidotus from the Buzi River. Hoots were only observed in agonistic encounters. Hoot oscillograms showed a sinusoidal waveform, and the mean duration of this sound was similar in Incomati system fish (mean, 161 ms), Upper Zambezi fish (172 ms) and Buzi fish (103 and 145 ms for the two vocalising individuals). The mean frequency of the first hoot harmonic was higher in Incomati system fish (326 Hz) than in Upper Zambezi fish (245 Hz). Both growl and hoot occurred only in the presence of conspecifics, probably signalling the presence and condition of an opponent, territory owner or potential mate. This is the first evidence for (1) sound production and acoustical communication in another species and genus, M. macrolepidotus, from southern Africa to be (2) geographically differentiated.