Citation: Connolly, R. M., and N. J. Waltham. 2015. Spatial analysis of carbon isotopes reveals seagrass contribution to fishery food web. Ecosphere 6(9):148. http://dx.doi.org/10.1890/ES14-00243.1Abstract. Despite the widespread use of carbon stable isotopes to distinguish among potential energy pathways in food webs, their usefulness is limited where potential basal carbon sources are numerous and diverse. We measured carbon isotope values of the major fisheries species, the mostly carnivorous Scylla serrata (giant mud crab), and potential basal, autotrophic sources supporting the food web. Conventional mixing modelling of autotroph and crab isotope data could not differentiate contributions from different sources. Pooling of modelled contributions from sources with similar isotope values indicated a role for organic matter from seagrass meadows or saltmarshes, but still did not define contributions well. Crab isotope data from a subsequent, spatially explicit survey of 14 sites, selected to represent different distances from key habitats, were analyzed using multiple regression. Crab isotope values showed a significant relationship with distance from seagrass (R 2 ¼ 0.87), but not with distance from mangroves or saltmarsh grass. Alongside seagrass meadows, crabs had very enriched isotope values, demonstrating their reliance on sources with enriched isotope values (seagrass and algae epiphytic on seagrass, 65-90% of their energy intake). At the site furthest from seagrass (21 km), crabs assimilated carbon primarily from depleted sources such as mangroves and terrestrial organic matter from coastal catchments (70-85%). Explicit spatial analysis of isotope data following a comprehensive survey revealed energy pathways not evident in conventional analyses.