Echolocation signal design in nine syntopic vespertilionid bats from the Malaysian rain forest understorey was studied. Four species of Kerivoula, two species of Phoniscus (Kerivoulinae) and three species of Murina (Murininae) all emitted calls that are typical of species that glean insects from surfaces: broadband, frequency-modulated (FM) calls of low intensity and short duration. However, calls were highly distinctive in the use of very large bandwidths (range: 89–123 kHz) and extremely high frequencies (start frequency 152–180 kHz; end frequency 43–86 kHz). Furthermore, calls were produced in groups of 2–15 at very high pulse repetition rates (37–105 Hz). The functional significance of these characteristics with respect to foraging strategy is discussed. Large signal bandwidths facilitate highly accurate target localization in terms of both range and angle estimation and can thus be interpreted as an adaptation to foraging in the highly cluttered environment of the forest understorey. The use of high frequencies so far in excess of those seen in other FM gleaning bats of the same size is less easily explained, but may represent a mechanism by which these species distinguish prey items using echolocation alone, without recourse to visual or auditory cues. Species exhibited differences in echolocation parameters, particularly the end frequency and the number of calls per group of calls. The two subfamilies differed from each other in multivariate space derived from echolocation parameters; calls of the Murininae were of lower frequency than the Kerivoulinae and were typically produced in smaller groups or singly. Within the subfamilies there was considerable overlap between species of Murina, but the Kerivoula spp. were clearly distinct from one another but not from the two Phoniscus spp.