|During 2002 and 2003 samples, collected every five miles along the Ohio River, were analyzed for total cultivable bacteria, ciprofloxacin resistant bacteria, erythromycin resistant bacteria, tetracycline resistant bacteria, and fecal coliforms. During 2004 samples were analyzed for total cultivable bacteria, ciprofloxacin resistant bacteria, sulfamethizole resistant bacteria, tetracycline resistant bacteria, Virginiamycin resistant bacteria, total coliforms, and Escherichia coli. The objectives of this study were to systematically collect data on fecal coliforms, E. coli, and antibiotic resistant bacteria in the Ohio River and its major tributaries; to determine if antibiotic resistance populations are correlated to each other or to coliforms; and to investigate antibiotic resistance patterns, spatially, over time, along the Ohio River. Data from 2002 and 2003 suggest that ciprofloxacin resistant, erythromycin resistant, tetracycline resistant, and coliform bacteria represented significantly different (P< 0.05) populations. Data from 2004 showed ciprofloxacin resistant, sulfamethizole resistant, tetracycline resistant, Virginiamycin resistant, total coliform bacteria and E. coli represented significantly different (P< 0.05) bacterial populations. This lack of correlation between resistant bacterial populations and coliform bacteria suggests antibiotic resistance in the Ohio River stems from sources other than fecal contamination. It can also be concluded from this study that antibiotic resistant populations and coliform bacteria are present in higher numbers when river flows are above harmonic mean flow.