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Little is known about the association of ambient ozone with ovarian reserve. Based on a retrospective cohort study of 6008 women who attended a fertility center in Hubei, China, during 2018–2021, we estimated ozone exposure levels by calculating averages during the development of follicles (2-month [W1], 4-month [W2], 6-month [W3]) and 1-year before measurement (W4) according to Tracking Air Pollution in China database. We used multivariate logistic regression and linear regression models to investigate association of ozone exposure with anti-müllerian hormone (AMH), the preferred indicator of ovarian reserve. Each 10 μg/m3 increases in ozone were associated with 2.34% (0.68%, 3.97%), 2.08% (0.10%, 4.01%), 4.20% (1.67%, 6.67%), and 8.91% (5.79%, 11.93%) decreased AMH levels during W1-W4; AMH levels decreased by 15.85%, 11.90%, 16.92% in the fourth quartile during W1, W3, and W4 when comparing the extreme quartile, with significant exposure-response relationships during W4 (P < 0.05). Ozone exposure during W1 was positively associated with low AMH. Additionally, we detected significant effect modification by age, body mass index, and temperature in ozone-associated decreased AMH levels. Our findings highlight the potential adverse impact of ozone pollution on female ovarian reserve, especially during the secondary to small antral follicle stage and 1-year before measurement. [Display omitted] • Ambient ozone exposure was associated with diminished ovarian reserve. • Secondary to small antral follicle stage might be the susceptible exposure window. • Even below the current AQS, AMH levels decreased with increasing ozone exposure. • Age, BMI, and ambient temperature might modify the observed associations. [ABSTRACT FROM AUTHOR]