Prior studies have established a significant correlation between the DNA fragmentation index (DFI) and infertility. Additionally, certain investigations suggest that environmental exposure may serve as an etiological factor impacting semen quality. This study aimed to explore the impact of season, ambient temperature, and weather extremes on the DFI of sperm, along with other relevant parameters. Furthermore, it sought to assess how ambient temperature affects the DFI of sperm and other semen parameters in populations with varying BMI values. Additionally, the study analyzed the transient indirect effect of DFI on sperm parameters. This retrospective study analyzed semen samples from 11,877 men, selected based on female factor considerations, spanning from January 2016 to December 2021. Participants were grouped according to the season of semen collection. The results showed that samples collected in summer had a lower semen volume and sperm motility, while those collected in autumn had a lower DFI. We analyzed the exposure-response ratio between environmental exposure temperature and semen parameters using a generalized additive model. Results showed that the curve of the exposure-response relationship was U-shaped or inverted U-shaped; when the air temperature exposure was below the threshold, for each degree of temperature increase, the total sperm motility, sperm concentration, and progressive motility increased by 0.16 %, 0.29 × 10 (Levine, 1999)/ml and 0.14 %, respectively, while the DFI and inactivity rate decreased by 0.078 % and 0.15 %, respectively. When the air temperature exposure exceeded the threshold, for each degree of temperature increase, the sperm concentration, total sperm motility, semen volume and progressive motility decreased by 0.42 × 10 (Levine, 1999)/ml, 0.11 %, 0.0078 ml and 0.15 %, respectively, while the DFI and inactivity rate increased by 0.13 % and 0.12 %, respectively. Extremely cold weather during spermatogenesis was positively correlated with DFI, and extremely hot weather was negatively correlated with sperm motility. Subgroup analysis revealed that individuals classified as overweight / obese exhibited more pronounced changes in sperm parameters and the DFI in response to variations in environmental exposure temperature compared to those with a normal BMI. In the analysis of the relationship between DFI and sperm parameters, the results showed an inverted U-shape relationship between DFI and semen volume, and a negative correlation between DFI and sperm concentration and sperm motility. And we found that ambient temperature affects sperm parameters through DFI at low as well as average temperatures, whereas at high temperatures this indirect effect is no longer present.