The research aimed to use chemical, geochemical, and ecotoxicity indices to assess the heavy metals content in soils with different degrees of exposure to human pressure. The research was conducted in southern Poland, in the Malopolska (Little Poland) province. All metal contents exceeded geochemical background levels. The highest values of the Igeo index were found for cadmium and were 10.05 (grasslands), 9.31 (forest), and 5.54 (arable lands), indicating extreme soil pollution (class 6) with this metal. Mean integrated pollution index (IPI) values, depending on the kind of use, amounted to 3.4 for arable lands, 4.9 for forests, and 6.6 for grasslands. These values are indicative of a high level of soil pollution in arable lands and an extremely high level of soil pollution in grasslands and forests. Depending on the type of soil use, Vibrio fischeri luminescence inhibition was from -33 to 59% (arable lands), from -48 to 78% (grasslands), and from 0 to 88% (forest). Significantly the highest toxicity was found in soils collected from forest grounds.