Drug-metabolizing cytochrome P450 (CYP) enzymes are especially important in wild animals as they are directly exposed to environmental pollutants and bioactive molecules of plants. Our main goal was to monitor the activity of certain CYP enzymes in wild boar compared to domestic swine, and to assess various modulatory factors of xenobiotic biotransformation in wild boar. Liver and intestinal mucosa (duodenum, jejunum, ileum, caecum) samples were collected from 49 hunted free-range wild boars and 15 wild boar fetuses; domestic pig samples (n = 40) were gained from a slaughter house. Specific activity of CYP1A2, CYP2C9, and CYP3A4 enzymes was assessed by luminometric assays. The activity of hepatic CYP1A2 and CYP3A4 enzymes was significantly higher in wild boars than in domestic pigs, while CYP2C9-mediated hepatic metabolism was significantly less intense in wild boars than in pigs. Certain modulatory factors (sex, sexual maturation, and season) were also confirmed in wild boars. The activity of all investigated intestinal CYP enzymes remained under detection level in each gut section in both species. Hepatic CYP2C9 and CYP3A4 enzymes were measurable in wild boar fetuses, but their activity was remarkably lower than in adults. The described interspecies differences might be explained with the altered exposure of wild and domesticated animals to specific CYP modulators. As CYP enzymes in wild boars can be highly influenced by environmental pollutants, following further studies, they may serve as ecotoxicological markers of agricultural or industrial toxicants. Investigating CYP-related drug metabolism in wildlife species can clarify some toxicokinetic interactions, thus having huge importance in the production of safe game meat.