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Discharge of saline groundwater from Eucalyptus forests on a semi-arid floodplain was directly determined by first measuring transpiration rates from the forests, and then partitioning the transpiration flux into groundwater discharge and soil water depletion. This partitioning was achieved by identifying the source of the transpired water with naturally occurring stable isotopes of water. Transpiration rates were low, being about 0.3 mm day-1 from three E. largiflorens sites and up to 2 mm day-1 from an E. camaldulensis site. There was no significant variation in transpiration across seasons, indicating that transpiration was limited by environmental factors other than evaporative demand. Despite its salinity (electrical conductivities of 11-33 dS m-1), the groundwater was used by the forests at all sites and all times, and made up 100% of transpiration in more than half of the measurements, and 40-80% in the remainder. There was some consistency in water uptake patterns. E. camaldulensis tended to take up shallow soil water and groundwater simultaneously, as did trees at one of the E. largiflorens sites. At the driest sampling time, however, groundwater was the only source of water for trees at both of these sites. Trees at the remaining two E. largiflorens sites generally relied solely on the groundwater. The tree water source results indicate that groundwater discharge fluxes were between 40 and 100% of the transpiration fluxes at these sites. These groundwater discharge fluxes were small in terms of regional groundwater balances, but would be important in the salinisation of the soils. Additionally, uptake of water from the soil profile by the trees substantially increased groundwater discharge compared with discharge from the soils had they been bare of vegetation.