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Tooth enamel apatite carbonate carbon and oxygen isotope ratios of modern kangaroos (Macropus spp.) collected on a 900-km latitudinal transect spanning a C-3-C-4 transition zone were analysed to create a reference set for palaeoenvironmental reconstruction in southern Australia. The carbon isotope composition of enamel carbonate reflects the proportional intake of C-3 and C-4 vegetation, and its oxygen isotope composition reflects that of ingested water. Tooth enamel forms incrementally, recording dietary and environmental changes during mineralisation. Analyses show only weak correlations between climate records and latitudinal changes in delta C-13 and delta O-18. No species achieved the delta C-13 values (similar to-1.0 aEuro degrees) expected for 100 % C-4 grazing diets; kangaroos at low latitudes that are classified as feeding primarily on C-4 grasses (grazers) have delta C-13 of up to -3.5 aEuro degrees. In these areas, delta C-13 below -12 aEuro degrees suggests a 100 % C-3 grass and/or leafy plant (browse) diet while animals from higher latitude have lower delta C-13. Animals from semi-arid areas have delta O-18 of 34-40 aEuro degrees, while grazers from temperate areas have lower values (similar to 28-30 aEuro degrees). Three patterns with implications for palaeoenvironmental reconstruction emerge: (1) all species in semi-arid areas regularly browse to supplement limited grass resources; (2) all species within an environmental zone have similar carbon and oxygen isotope compositions, meaning data from different kangaroo species can be pooled for palaeoenvironmental investigations; (3) relatively small regional environmental differences can be distinguished when delta C-13 and delta O-18 data are used together. These data demonstrate that diet-isotope and climate-isotope relationships should be evaluated in modern ecosystems before application to the regional fossil record.