Freshwater habitat is a natural reservoir for antimicrobial resistance (AMR). AMR poses serious human, animal, and environmental public health threats. This study aimed to evaluate the physicochemical and microbiological quality of five selected rivers (Apitipiti 1, Apitipiti 2, Apitipiti 3, Sogidi, and Aba Apa Akinmorin) in Oyo town, Nigeria, as well as the antibiotic resistance pattern of isolated bacterial species, using conventional methods. Most physicochemical parameters were within WHO and NIS permissible limits. Pearson’s correlation matrix indicated that there were significant (p < 0.05) interactions among pH, electrical conductivity, temperature, sulphate and chloride salts, and BOD and COD. A total of thirty-two (32) bacterial species were isolated and identified as: Aeromonas (9), Bacillus (2), Corynebacterium (13), Lactobacillus (1), Pseudomonas (2), Staphylococcus (4), and Streptococcus (1). Of the rivers, Sogidi had the highest microbial load (6.36 log CFU/mL) while Apititipiti 1 had the lowest (5.76 log CFU/mL). With regard to antibiotic sensitivity, 81.8% were multidrug-resistant, with Corynebacterium kutscheri and Aeromonas spp. isolated from Apitipiti 2 and Aba Apa Akinmorin rivers, respectively, exhibiting a relatively high antibiotic resistance of 90.9%. This study reveals that these rivers may be unfit for consumption as multidrug-resistant bacteria of public health risk were associated with them.