Huanglongbing (HLB) is a devastating bacterial disease associated with 'Candidatus Liberibacter asiaticus'. The location of the pathogen within the vasculature of the tree has left growers with limited options for the effective management of the disease. Trunk injection is a crop protection technique that applies therapeutics directly into the xylem of woody tree species and allows for their systemic uptake and transport, which may provide more effective management of vascular diseases such as HLB. In this study, mature 'Valencia' and 'Hamlin' sweet orange (Citrus sinensis) and 'Duncan' grapefruit (C. paradisi) trees were injected with oxytetracycline (OTC) in the spring and/or fall to evaluate the effects of injection timing and response to injection. In addition to seasonal evaluations of tree health and bacterial titer, preharvest fruit drop, yield, and fruit quality were measured at harvest to determine the effects of OTC injection. The benefits associated with injection included a reduction in fruit drop, an increase in fruit yield and fruit size, and improvements in juice quality. However, results varied due to the timing of injection and were not consistent across all three varieties. Residue analysis at different time points after injection suggests that trunk injection effectively delivers therapeutics to mature citrus trees. This study provides fundamental information on the short-term benefits associated with trunk injection of OTC for HLB management in citrus groves. The potential for use of trunk injection at the commercial scale and the possible risks are discussed.