In this study, detection of a possible relationship of some trace element levels of copper (Cu), zinc (Zn), and iron (Fe), toxic heavy metals, cadmium (Cd) and lead (Pb), and antioxidant functions in cattle, which are living near the trunk roads for at least 3 years, was aimed. Totally, 70 cattle, which were in different herds, were used. Fifty-four cattle (3-13 years), which were living near the trunk roads at about 0-500 m were used for the experiment group. Also, 16 cattle (3-12 years), which were living far from trunk roads at about 2-3 km were used for the control groups. Blood samples were collected and Cd, Pb, Cu, Zn, Fe levels, lipid peroxidation product malondialdehyde (MDA) and antioxidative defense system indicators, erythrocyte superoxide dismutase (SOD) activity, catalase (CAT) activity, and plasma ceruloplasmin (CP) level were measured. Cd and Pb levels of the experimental animals were determined to be higher than the control group (P<0.01 and P<0.001, respectively). However, Zn level of the experiment group was found lower than the control animals (P<0.001). MDA increased about three times and CP increased about two times more than the control group (P<0.001). Significantly correlations were observed among Pb, MDA and CP levels. Also, important correlations were found between Fe and CP, also Cu and MDA levels. Besides, MDA and CAT levels of the animals increased with CP and SOD together, respectively. At the end of the study, Cd and Pb accumulation was observed in cattle, which were living near the road with intensive traffic for at least three years when compared with the control group. In addition, the results showed significant interactions among toxic heavy metals, physiological trace elements, enzymatic and non-enzymatic antioxidants and the lipid peroxidation.