Pruning of live branches is a management option to enhance wood quality in plantation trees. It may also alter whole-tree water use, but little is known about the extent and duration of changes in transpiration. In this study, sap flow sensors were used to measure transpiration for 14 days prior to, and 75 days following the removal, through pruning, of the lower 50% of the live-crown length of 10–11 m tall four-year old Eucalyptus pilularis Sm. and E. cloeziana F. Muell. trees. Pruning had no effect on stem growth, sapwood water content or radial pattern of sap velocity in either species. Pruning reduced mean daily water use by 39% in E. pilularis and 59% in E. cloeziana during the first eight days after pruning. Thirty six days after pruning there were no longer any significant differences in transpiration rates between pruned and unpruned trees in either species. Our results show that pruning of live branches had only a short-term effect on whole-tree transpiration in these sub-tropical eucalypt species.