Background:As a complementary means to urban public transit systems, public bike-sharing provides a green and active mode of sustainable mobility, while reducing carbon-dioxide emissions and promoting health. There has been increasing interest in factors affecting bike-sharing usage, but little is known about the effect of ambient air pollution. Method:To assess the short-term impact of daily exposure to multiple air pollutants (PM2.5, PM10, NO2, and O3) on the public bike-sharing system (PBS) usage in Seoul, South Korea (2018–2021), we applied a quasi-Poisson generalized linear model combined with a distributed lag nonlinear model (DLNM). The model was adjusted for day of the week, holiday, temperature, relative humidity, and long-term trend. We also conducted stratification analyses to examine the potential effect modification by age group, seasonality, and COVID-19. Results:We found that there was a negative association between daily ambient air pollution and the PBS usage level at a single lag day 1 (i.e., air quality a day before the event) across all four pollutants. Our results suggest that days with high levels of air pollutants (at 95th percentile) are associated with a 0.91% (0.86% to 0.96%) for PM2.5, 0.89% (0.85% to 0.94%) for PM10, 0.87% (0.82% to 0.91%) for O3, and 0.92% (0.87% to 0.98%) for NO2, reduction in cycling behavior in the next day compared to days with low levels of pollutants (at 25th percentile). No evidence of effect modification was found by seasonality, age nor the COVID-19 pandemic for any of the four pollutants. Conclusions:Our findings suggest that high concentrations of ambient air pollution are associated with decreased rates of PBS usage on the subsequent day regardless of the type of air pollutant measured.