The objectives of the present study were first to determine the residual contents of total aflatoxins (AFTs), lead (Pb), and cadmium (Cd) in the edible tissues of the cattle reared in Al-Ahsa, Saudi Arabia. Al-Ahsa is the largest governorate in the Eastern Province of Saudi Arabia. The two main economic activities of Al-Ahsa are oil production (industrial) and agriculture. Besides, dietary intake and possible health risks for Saudi population were further calculated. In order to establish potential molecular biomarkers for xenobiotic exposure in cattle, the mRNA expression of xenobiotic-metabolizing enzymes (XMEs) including cytochrome P450 (CYP) 1A1, NAD(P)H dehydrogenase [quinone] 1 (NQO1), metallothionein (MT) 1A, and heat shock protein (HSP) 70 was investigated in the different tissues of the cattle. The tested XMEs were selected because of their specific roles in the metabolism and detoxification of AFTs, Pb, and Cd. The obtained results revealed that the liver had significantly the highest AFT content, while all examined muscle samples had no AFT residues. Consumption of the bovine liver and kidneys represents the highest source for the dietary exposure to total AFTs (0.05–0.98 μg/kg/day), Pb (0.06–0.19 mg/kg/day), and Cd (0.08–0.19 mg/kg/day) among the examined tissues. Therefore, excessive intake of such organs might pose a public health concern, particularly among children. Significant upregulation of mRNA expressions of CYP1A1, NQO1, MT1A, and HSP70 was observed in the different tissues of the cattle in comparison with the muscle. This upregulation had significant positive correlation with the accumulated AFTs, Pb, and Cd. This indicates the possible use of CYP1A1, NQO1, MT1A, and HSP70 as potential biomarkers for the exposure of the cattle to AFTs, Pb, and Cd.