Walking Football is an adaptation of Association Football, played primarily by middle-aged and older adults, with rule changes to enhance accessibility. Sport participation poses injury risks, yet the risk of injury in Walking Football has not been established. This study quantifies injury incidence, estimates injury burden, and examines injury characteristics. The prospective observational study recorded injuries during The FA Walking Football Cup 2024, featuring 84 teams in three categories: Women's 40 years+, Mixed Gender 50 years+, and Mixed Gender 60 years+. Time-loss injuries (≥1 day estimated absence), and medical attention injuries (on-field attention only) were measured along with match exposure (total playing-hours). Injury incidence and estimated injury burden per 1000 player-hours, and injury characteristics, were analysed. Across all categories, 42 match injuries occurred over 850.6 h of exposure, with 8 time-loss (19%) and 34 medical attention injuries (81%). The overall injury incidence was 49.4 injuries per 1000 h, with time-loss injury incidence at 9.4 and medical attention injury incidence at 40.0 injuries per 1000 h. The total estimated injury burden was 156.4 days absent per 1000 h. Injuries were evenly distributed between non-contact (52%) and contact mechanisms (48%), with tackles representing 45% of cases and running, explicitly prohibited in Walking Football, accounting for 12%. Overall, Walking Football has a low injury risk, with most injuries classified as non-time-loss. However, the prevalence of contact-related injuries suggests stricter enforcement of minimal contact rules could further reduce injury incidence. Safety strategies should prioritise reducing tackle-related injuries and minimising running to ensure player welfare.