Intensive cattle breeding’s environmental challenges are prompting shifts to extensive, pasture-based systems, influencing nutrient and pollutant uptake. PTEs are essential and non-essential elements, regularly found in the environment and organisms, and in which unbalances lead to health issues. Hair analysis, a non-invasive method, provides retrospective PTE exposure evaluation. This study aims to understand exposure and species-specific accumulation patterns of PTEs in cattle and roe deer sharing pastures in Northern Italy using the hair analysis. Aluminum, As, Cd, Cr, Ni, Pb, Cu, Mg, Fe, and Zn were quantified through the use of ICP-OES. Findings show As levels significantly higher in roe deer due to their selective feeding, while Cd and Pb levels align with other studies. Essential elements like Cu, Fe, and Zn are lower in cattle, possibly due to diet differences. Higher Cr and Ni levels in cattle suggest contamination or physiological differences in accumulation patterns. In conclusion, hair analysis is valuable for monitoring environmental PTE exposure, highlighting significant interspecies differences and the potential of both animals as bioindicators in shared grazing areas.