The concentration of Al, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Sb, Se, V and Zn was measured in the aquatic bryophyte Rhynchostegium riparioides after a 4-week transplantation at 83 stations in seven main watercourses and three smaller tributaries of the basin of the River Bacchiglione (NE Italy). The study, carried out in 9 months, aimed at establishing a database for statistical analyses to define a novel index of water quality, specifically calibrated for moss bags, and suitable for bio-monitoring purposes where autochthonous bryophytes are not available. The background level was estimated for each element, the ratio between sample and background concentration was adopted as measure of environmental alteration and a five-class interpretation scale was defined. The 73.8% of measured concentrations reflected a condition of naturality, 16.6% suspect of alteration, 7.6% sure alteration, 1.6% severe alteration and <0.5% extreme alteration. The condition of global environmental alteration (all contaminants) at each station was also defined. Forty-four stations presented absence or suspect of alteration, 39 sure alteration for one or more elements. Bivariate and multivariate analyses revealed (1) highly significant correlations (p < 0.001) between the concentrations of Co–Mn, Sb–Zn, and Fe–V, depending on both natural and human factors, (2) chemical patterns leading to galvanic industries, metallurgy or urban sewages, in some case with a geographical distribution and (3) a gradient of environmental alteration from the Pre-Alps to the lowland. Coloured maps indicate both point sources and widespread environmental alteration, highlighting not only industrial–urban areas but also some apparently undisturbed sites