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The aim of the study was to determine the effects of land use management on changes in the fecal contamination of water in the Łyna River, one of the main lowland watercourses in the southern watershed of the Baltic Sea (northern Poland). A total of 120 water samples were collected in different seasons of 2011 and 2012 at 15 sites where the river intersected forest (FA), agricultural (AA), and urbanized (UA) areas. Fecal indicator bacteria (FIB), the counts of Enterobacteriaceae and Escherichia coli, total bacterial counts (TBCs), and domain Bacteria (EUB338) were determined by culture-dependent and culture-independent methods. Temperature, pH, chemical oxygen demand, dissolved oxygen, total dissolved solids, ammonia nitrogen, nitrite nitrogen, nitrate nitrogen, orthophosphate, and total phosphorus were also determined. The lowest bacterial counts were noted in water samples collected in FA, and the highest in samples collected in UA. Statistically significant differences were determined between bacterial populations across the analyzed land use types and in different sampling seasons. Significant correlations were also observed between the populations of FIB and physicochemical parameters. The results indicate that land use type influenced FIB concentrations in river water. The combined use of conventional and molecular methods improves the accuracy of fecal contamination analyses in river ecosystems.