In autumn 2005, the site-specific variability of cytosolic metal and protein concentrations in gills of European chub from the Sava River could be mostly associated with gill mass variability. In spring 2006, the correlations of metals and proteins with gill mass were mainly nonsignificant, and their site-specific variability could be presumably associated with river water pollution. The spring cytosolic concentrations of Zn, Cu, and Mn did not differ significantly between the sites (medians: 8.37-11.34 microg ml(-1), 68.2-86.2 ng ml(-1), and 55.9-68.6 ng ml(-1), respectively). Increased cytosolic Cd concentrations were obtained at Oborovo and Lukavec Posavski (median: 4.01 ng ml(-1))-the sites influenced by pollution sources from two major urban areas-compared with the remaining sampling sites (median: 1.93 ng ml(-1)). Cytosolic Fe concentrations were almost two times higher at Jasenovac (median: 11.98 microg ml(-1)) compared with the concentrations at Sava-Zagreb (median: 6.72 microg ml(-1)). Labile Fe concentrations measured in river water with passive samplers indicated that cytosolic Fe concentrations in the spring possibly reflected water-borne Fe uptake. The spring cytosolic protein concentrations decreased from upstream (Otok Samoborski: 27.2 +/- 5.6 mg ml(-1)) toward the downstream sites (Lukavec Posavski: 14.8 +/- 2.8 mg ml(-1)), possibly because of the influence of organic pollution and water toxicity. The spring period seems to be a more appropriate season for the assessment of the river water pollution if chub gills are used as the target organ.