In aquatic systems, the bioavailability of an element to microorganisms is greatly influenced by its chemical speciation. The goal of this work was to assess metal toxicity to a green algae (Pseudokirchneriella subcapitata) and a bacterium (Vibrio fisheri) as a function of size fractionation and chemical speciation (using the program MINTEQA2) in contaminated water of the Upper Vistula River. Water samples were collected at 1 reference site, 4 polluted sites and one polluted site on Vistula's main tributary, the Przemsza River. Toxicity measurements were performed on unfiltered samples and, total dissolved (<1.2μm), and truly dissolved (<1 kDa) fractions. Trace metal (Cd, Co, Cr, Cu, Mn, Pb, Zn) concentrations were measured in these samples and also in the colloidal fraction (1 kDa-1.2μm). At the reference site, the low metal concentrations were in agreement with the absence of measurable toxicity. In the polluted section of the river, free metal concentrations were largely below the potential toxic levels for bacteria, wich was in agreement with the absence of toxicity. Although Zn²⁺was at potentially toxic-level concentrations in total dissolved and truly dissolved fractions in the polluted riverine section, toxicity for algae was observed, only in truly dissolved fractions from two stations. The absence of toxicity in most samples was related to metal association with particles and with low molecular weight ligands as well as the presence of organic ligands (phenol). The reason for toxic effects in two ultrafiltered samples is not clear, but may be related to the elimination of the colloidal organic fraction and thus the eradication of its protective effect occuring in natural samples.