The spatial and temporal distribution of heavy metals in water, sediment, fish and mussel of Dhaleswari River, Bangladesh were determined by atomic absorption spectrophotometer. In water, the concentrations of Ni, Pb, Cd, Cr and Cu varied seasonally and spatially from 5.47-9.74, 38.25-63.28, 5.29-8.20, 378.87-501.11 and 98.37-188.08 μg/L, respectively. The sediment also showed a spatial and temporal variation of Ni, Pb, Cd, Cr and Cu ranges from 135.02-231.44, 58.19-70.26, 2.11-4.14, 95.76-141.27 and 31.53-76.52 mg/kg, respectively. These variations are likely to be due to different collection spots with point and non-point sources and seasons. The concentrations of Cd, Cr and Cu were higher, while Ni and Pb were lower in water than those of sediment. In Trypauchen vagina, a bottom-living fish, the concentration of Ni, Pb, Cd, Cr and Cu varied seasonally from 6.35-9.56, 6.14-8.03, 0.51-0.73, 6.92-12.23 and 5.43-9.45 mg/kg, respectively. The concentrations of heavy metals in this fish were much lower than those of water and sediment. In Glossogobius giuris, also a bottom-living fish, the concentration of Ni, Pb, Cd, Cr and Cu varied seasonally from 4.75-10.17, 4.25-8.17, 0.61-0.71, 7.15-11.92, and 5.17-7.48 mg/kg, respectively, which were slightly lower (except Cd) than those of T. vagina. In Lamellidens marginalis, a freshwater bivalve, the concentration of Ni, Pb, Cd, Cr and Cu varied seasonally from 6.07-11.32, 7.03-59.21, 0.56-7.23, 9.38-501.11 and 7.55-183.87 mg/kg, respectively. The concentrations of all the heavy metals studied were much higher in mussel than those of fish that indicate the greater rate of bio-accumulation in mollusc.