The scale and patterns of movement and habitat use are primary considerations in the conservation and management of threatened species. Movement, activity and habitat use of the threatened two-spined blackfish Gadopsis bispinosus were assessed in a small upland reservoir in south-eastern Australia using manual and remote radio-telemetry. Movements and activity of two-spined blackfish (n = 19) were studied over a 28-day period and exhibited proportionately large directional crepuscular movement and activity with heightened activity continuing throughout the night (although movement was subdued). Two daily movement strategies were observed: movements from diurnal home-shelter habitats (predominantly rock) to macrophytes at night (14 individuals), and occupation of macrophytes during the entire diel period and restricted movement (five individuals). Daily movement strategies were fixed (not plastic) among all individuals, with one exception, for the duration of the study period. Rock, fallen timber and macrophytes were the most commonly used daytime shelter habitat (in order of reference). Although some information exists on movements and habitat use of this species and the congeneric river blackfish G. Marmoratus in lotic environments, we present the first study of movements and habitat use for either species in lentic environments. Given the occupation of lentic environments by this threatened species, the data presented in this study provide insight into the habitat requirements for this species, and offer opportunities for habitat enhancement in existing reservoirs within the species geographic range.