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The Impact of Same-Day Antiretroviral Therapy Initiation under the WHO Treat-All Policy.Rapid initiation of antiretroviral therapy (ART) is recommended for people living with HIV, with the option to start treatment on the day of diagnosis (same-day-ART). However, the effect of same-day-ART remains unknown in realistic public sector settings. We established a cohort of ≥16-year-old patients who initiated first-line ART under Treat-All in Nhlangano (Eswatini) between 2014-2016, either on the day of HIV care enrolment (same-day-ART) or 1–14 days thereafter (early-ART). Directed acyclic graphs, flexible parametric survival analysis and targeted maximum likelihood estimation (TMLE) were used to estimate the effect of same-day-ART initiation on the composite unfavourable treatment outcome (loss to follow-up;death;viral failure). Of 1328 patients, 839 (63.2%) initiated same-day ART. The adjusted hazard ratio of the unfavourable outcome was increased by 1.48 (95% CI:1.16–1.89) for same-day-ART compared with early-ART. TMLE suggested that after 1 year, 28.9% of patients would experience the unfavourable outcome under same-day-ART compared with 21.2% under early-ART (difference: 7.7%; 1.3–14.1%). This estimate was driven by loss to follow-up and varied over time, with a higher hazard during the first year after HIV care enrolment and a similar hazard thereafter. We found an increased risk with same-day-ART. A limitation was possible silent transfers that were not captured.