With urbanization and industrialization, air pollution is one of the known health hazards and a severe problem in human societies. This study assessed the relationship of SO2, O3, and ambient air pollution on hematologic factors, serum iron, and TIBC in a rat model. A total of 32 male Wistar rats were enrolled in the study. They were randomly divided into the four study groups (n = 8 in each) as follows: The control, SO2 group exposed to 10 ppm, ozone group (O3) exposed to 0.6 ± 0.1 ppm, and ambient air pollution (AAP) group for five weeks (3 h/day). One-way analysis of variance was used to With urbanization and industrialization, air pollution is one of the known health hazards and a severe problem in human societies. This study assessed the relationship of SO2, O3, and ambient air pollution on hematologic factors, serum iron, and TIBC in a rat model. A total of 32 male Wistar rats were enrolled in the study. They were randomly divided into the four study groups (n = 8 in each) as follows: The control, SO2 group exposed to 10 ppm, ozone group (O3) exposed to 0.6 ± 0.1 ppm, and ambient air pollution (AAP) group for five weeks (3 h day-1). One-way ANOVA (analysis of variance) was used to compare parameters. Significant changes in RBC (P=0.026), hemoglobin concentration (P=0.029), mean corpuscular volume (MCV) (P=0.011), mean corpuscular hemoglobin concentration (MCHC) (P<0.001), monocytes (P=0.002), and basophils (P=0.022) were observed between control and AAP groups. The other parameters showed an insignificant difference for these two groups' comparison. In the of leucocytes’ differential counts, only basophils and monocytes were statistically high in the AAP group compared to the control group (P=0.002 and P=0.022, respectively). In terms of serum iron, the differences between the control and SO2 groups (P =0.008), or between the groups of O3 and APP (p=0.028) were statistically significant. The observed hematological and biochemical changes indicated the toxic effects of ambient air particles. Further epidemiological studies are necessary to investigate the impact of other air pollutants like NO2, NO, and CO on hematological and biochemical parameters. Manuscript profile