Understanding the accumulation of potentially toxic elements (PTEs) across habitats is essential for evaluating ecological and human health risks. This study assessed the concentrations of ten PTEs (As, Cd, Co, Cr, Cu, Hg, Ni, Pb, Zn, and Mn) in soils and substrates, four habitat types (agricultural land, grassland, forest, and wetland), and in selected plants and wild mushrooms. The study encompassed the determination of total and mobile forms of these elements, thereby facilitating a comprehensive assessment of the monitored elements in the soil/substrate. The results provided insights into the state of soil contamination, which is crucial for the evaluation of potential environmental and health risks. The spatial distribution of the studied elements in the soil/substrate and plant material was visualised using maps generated through spatial interpolation. The study revealed extremely high concentrations of mercury and other contaminants, particularly in the vicinity of the emission source (former ore processing plant in Rudňany). The highest recorded Hg concentration was 411.13 mg kg (dry matter) in a forest substrate sample collected near the emission source. Cadmium also exceeded the limit value set for agricultural soil. The forest substrate exhibited the highest concentrations of the studied elements compared to other habitat types. The results of the bioaccumulation factor values indicated that the studied plants do not accumulate the elements. Wild mushrooms accumulated substantially higher levels of Hg compared to plants. It was reflected by THQ and %PTWI values that exceeded safe thresholds in some localities, indicating potential non-carcinogenic health risks for frequent mushroom consumers. Based on the Pollution Load Index, the whole territory can be characterised as heavily polluted. Extreme pollution levels were identified within a 10-kilometre radius, primarily in the southeast and south of the emission source, as well as at distances of 10 to 12 kilometres west and in the eastern part of the study area.