IntroductionOnline adaptive radiotherapy (OART) utilizes daily imaging to identify changes in the patient's anatomy and generate a new treatment plan adapted to these changes for each fraction. The aim of this study was to determine the intra-fraction motion and planning target volume (PTV) margins required for an OART workflow on the Varian Ethos system. MethodSixty-five fractions from 13 previously treated OART patients were analyzed for this retrospective study. The prostate and seminal vesicles were contoured by a radiation oncologist on two cone beam computed tomography scans (CBCT) for each fraction; the initial CBCT at the start of the treatment session, and the verification CBCT immediately before beam-on. In part 1 of the study, PTVs of different sizes were defined on the initial CBCT, and the geometric overlap with the clinical target volume (CTV) on the verification CBCT was used to determine the optimal OART margin. This was performed with and without a patient re-alignment shift by registering the verification CBCT to the initial CBCT. In part 2 of the study, the margins determined in part 1 were used for simulated Ethos OART treatments on all 65 fractions. The resultant coverage to the CTV on the verification CBCT, was compared to an image guided radiotherapy (IGRT) workflow with 7mm margins. ResultsPart 1 of the study found, if a verification CBCT and shift is performed, a 4 mm margin on the prostate and 5 mm on the seminal vesicles resulted in 95% of the CTV covered by the PTV in more than 90% of fractions, and 98% of the CTV covered by the PTV in more than 80% of fractions. Part 2 of the study found when these margins were used in an Ethos OART workflow, they resulted in CTV coverage that was superior to an IGRT workflow with 7mm margins. ConclusionA 4 mm prostate margin and 5 mm seminal vesicles margin in an OART workflow with verification imaging are adequate to ensure coverage on the Varian Ethos system. Larger margins may be required if using an OART workflow without verification imaging.