The anthropogenic load of antibiotics in Morogoro Municipality water bodies was estimated using the ELISA technique. Commercially available enzyme-linked immunosorbent assay (ELISA) kits commonly used for detection of tetracyclines, sulfonamides and quinolones residues in meat, milk, eggs and honey were adopted for analysis of these antibiotics in surface waters. Thirty-three sampling points were selected, including two rivers, one dam and one wastewater treatment plant. Results showed that there were detectable levels of the selected antibiotics in all the sampling sites. In rivers there were slightly higher mean concentration of antibiotics in the downstream rivers in close vicinity to high human activities. High mean concentrations of antibiotics were also found in wastewater treatment plant. In rivers, the maximum mean concentrations detected were 7.74±2.45, 8.76±2.65 and 8.94±3.83μg/l quinolones, tetracyclines and sulfonamides, respectively. At Mindu Dam, mean concentrations were, 1.61±0.17, 4.84±1.23 and 3.65±1.37μg/l, quinolones, tetracyclines and sulfonamides, respectively, while at Wastewater Treatment Plant mean concentrations were 31.55±11.23, 48.89±21.82 and 37.94±14.05μg/l quinolones, tetracyclines and sulfonamides, respectively. Differences in mean concentrations in Rivers, Dam and Wastewater Plant were attributed to the fact that in wastewater treatment plant antibiotic residues are continuously poured through human domestic and hospital waste, while at Mindu Dam and Rivers antibiotic residues are poured mainly through run-off during rainy season, hence high mean concentrations in Wastewater Treatment Plant than in rivers and dam. The presence of antibiotics residues in rivers, Mindu Dam and Wastewater Treatment Plant although in very low concentrations poses risks to population who take antibiotics at low doses. This can be through drinking water and consumption of crops produced through irrigation using water from these sources; this can lead to acceleration of antibiotic resistance through selection pressure; also can harm organisms in water ecosystem in long time exposure