Investigating resource partitioning among mobile marine predators such as cetaceans is challenging. Here we integrate multiple methodologies (analyses of habitat use, stable isotopes and trace elements) to assess ecological niche partitioning amongst two genetically divergent sympatric subpopulations (North and South) of Indo-Pacific bottlenose dolphins (Tursiops aduncus) in Moreton Bay, Australia. Comparisons of the mean locations (latitude, longitude) and environmental variables (distance from sandbanks, distance from shore and water depth) observed at sightings of biopsy-sampled individuals indicated that the North subpopulation occurred in the northwestern bay in significantly deeper water than the South subpopulation, which was found in southeastern nearshore waters and closer to sandbanks. Ratios of stable carbon and nitrogen isotopes in skin samples suggested that North dolphins foraged on higher trophic level prey in relatively more pelagic, offshore habitats, while South dolphins foraged on lower trophic prey in more nearshore, demersal and/or benthic habitats. Habitat partitioning was also reflected in higher blubber concentrations of most of the 13 measured trace elements, in particular lead, in the coastal South compared to the more pelagic North dolphins. These findings indicate that genetic subpopulations of bottlenose dolphins in Moreton Bay are adapted to different niches.