We report results from an evaluation of the levels of heavy metals, i.e., copper (Cu), cadmium (Cd), lead (Pb), nickel (Ni), manganese (Mn), chromium (Cr), and iron (Fe) in sediment and tilapia fish samples from a wide stretch of the Kafue river of Zambia. In sediment samples, the highest Pb and Fe concentrations were recorded at Hippo Dam, i.e., 36.2 ± 0.1 mg/kg dw and 733 ± 37 mg/kg dw at Kafue Town, respectively. Other notably high metal concentrations in sediment were Cr at Kafue Bridge (42.5 ± 0.1 mg/kg dw [dw]), Cu at Mpongwe (233 ± 5 mg/kg dw), and Mn at Kafue Town (133 ± 1 mg/kg dw); it was highest at Ithezi Tezhi Dam at 166 ± 1 mg/kg d. Three fish species, i.e., three-spot bream Tilapia andersonii, red-breasted bream T. rendalli, and nile tilapia Oreochromis niloticus were evaluated for levels of the seven metals. The concentrations of the metals in these fish species afforded estimation of the biota sediment-accumulation factor, which is the ratio of the concentration of the metal in liver to that in the sediment. The coefficients of condition (K) values, which give an indication of the health of the fish, were also estimated. The K values ranged from 2.5 ± 0.5 to 5.1 ± 0.6 in all of the three fish species. Partial least squares analysis showed that heavy metals are generally sequestered evenly in all of the parts of all of the three fish species except for elevated levels of Mn, Cd, and Pb in the liver samples.