Microbially produced extracellular polymeric substances (EPS) have been linked with many important ecological functions in natural sediments; yet, most information has been derived from marine systems. The present paper is the first comprehensive study on EPS (i.e., carbohydrates and proteins) dynamics in riverine sediments addressing spatial (six reservoirs and four groyne fields across three European rivers), temporal (all seasons in 2003-2005), and vertical (over a 50-cm sediment depth transect) pattern. The variation in hydrodynamic regime found in the reservoirs and groyne fields was reflected in the biomass and composition of the benthic microorganisms that produce EPS. The microphytobenthic communities consisted mainly of diatoms and a higher algal biomass (up to 248 microg g(-1) dry weight, DW) seemed to be indicative for higher amounts of secreted colloidal carbohydrates. Consequently, the model proposed by Underwood and Smith (1998) for the relation chlorophyll-colloidal carbohydrates was also applicable for upper riverine sediment layers. The close relation between algal biomass and bacterial cell counts (10(8)-10(9) cells g(-1) DW) supports the idea of bacterial use of the secreted EPS. However, the data also suggest a contribution to the EPS pool through bacterial secretion of proteins/extracellular enzymes and possibly carbohydrates. Over depth, the relationships between microorganisms and EPS became increasingly decoupled along with increasing ratios of bound (refractory) to colloidal (labile) EPS. These data suggest fresh production of polymeric substances in upper sediment layers and mainly accumulation of refractory, biodegraded material in deeper layers. The high contents of EPS colloidal and bound carbohydrates (0.1-1.8 and 1.3-6.7 mg g(-1) DW, respectively) and EPS proteins (0.4-12.9 mg g(-1) DW) at the freshwater study sites might indicate an important role in sediment ecology.