Wastewater from industrial and domestic sources flows into major bodies of water where people drink and bathe, resulting in a high incidence of severe water-borne infections. This study aims to compare the microbiological and physicochemical parameters of domestic and industrial wastewater in Ado-Odo LGA, Ogun State, Nigeria. Microorganisms were isolated from collected wastewater samples and were phenotypically identified using standard microbiological methods. Antimicrobial Sensitivity testing was carried out using Kirby Bauer`s disk diffusion technique. The physicochemical analysis of the water samples was also determined using standard analytical methods. A total of nineteen microorganisms were isolated from the samples collected with the predominant organism isolated from the two sources being Escherichia coli. The microorganisms isolated from the industrial wastewater sample were Citrobacter freundii (37.5%), Escherichia coli (37.5%), and Proteus vulgaris (25.0%) while those isolated from the domestic wastewater sample were Klebsiella oxytoca (18.18%), Escherichia coli (45.45%), Proteus vulgaris (9.09%), Staphylococcus saprophyticus (9.09%) and Staphylococcus aureus (18.18%). The physicochemical analysis conducted indicated that there were lower levels of impurities in industrial wastewater pH (4.1 ± 0.14), Electrical conductivity (479 ± 1.41), Total Dissolved Solids (223.5 ± 3.53), and Chloride content (168.12) compared to the domestic wastewater with pH (8.05 ± 0.07), Electrical Conductivity (1928± 4.24), Total Dissolved Solids (765.5 ± 0.70) and Chloride content (238.224) which suggests that industrial wastewater has been subjected to series of chemical and biological processes. The presence of Escherichia coli in higher levels in domestic wastewater indicates fecal contamination which is a major public health challenge since this wastewater leaks into various water bodies.