© 2015 Elsevier B.V. The inevitable development of resistance has sunk the great success achieved in the discovery of antimicrobial agents and dashed the hope of man in the recovery from infections and illnesses, as diseases and disease agents that were once thought to be controlled by antimicrobials are now re-emerging in new leagues resistance to therapy. A total of 300 PCR confirmed Escherichia coli isolates recovered from different river sources in Osun State, Nigeria were evaluated for their antibiogram profiling by the disc diffusion method and the resistant isolates were further profiled for their genotypic antimicrobial resistance determinants by polymerase chain reaction assays. Among the 20 antimicrobials selected from 10 families, resistance among sulfonamides, β-lactams and tetracyclines were found to be most frequent than phenicols and aminoglycosides with a noticeable increase in the number of multi-drug resistance ranging from three to nine antimicrobials. A total of 19 resistance determinants were assessed with their prevalence and distributions obtained as follows; [sulfonamides sulI (8%), sulII (41%)], [β-lactams; ampC 22%; bla<inf>TEM,</inf> (21%), and bla<inf>Z</inf> (18%),], [tetracyclines tetA (24%), tetB (23%), tetC (18%), tetD (78%), tetK (15%), and tetM, (10%)], [phenicols; catI (37%), catII (28%), and cmIA1 (19%)] and [aminoglycosides; aacC2 (8%), aphA1 (80%), aphA2 (80%), aadA (79%) and strA (38%)]. The Pearson chi-square exact test revealed many strong significant associations among ampC, bla<inf>TEM</inf>, bla<inf>Z</inf> and tetA genes with some determinants screened. The findings signify high increase in the prevalence of multidrug resistant E. coli isolates and resistance determinants indicating increased public health risks associated with the ingestion of waters from untreated sources. Hence, a necessity for safe water supply, provision of proper sanitation facilities and good surveillance programmes to monitor antimicrobial resistance patterns in water bodies.