Tetracycline resistance has been used as the key determinant to monitor resistance genes in natural environments such as rivers, lakes or seawater. The aim of this study was evaluate the frequency of tetA, tetB, tetC, tetD and tetE genes in 52 tetracycline-resistant Enterobacteriacea isolated from river water in the North East Black Sea Region of Turkey. In 52 tetracycline-resistant strains, resistance was mediated by tetA in eight (15.3%) strains, tetB in ten (19.2%) strains and both tetA and tetB in one (1.9%) strain. No tetC, tetD or tetE-mediated resistance was detected. In conclusion, the river water may be considered as a reservoir for the antibiotic resistance genes and the people living in this area may be under risk. To our knowledge, this is the first report for molecular characterization of tetracycline resistance in Enterobacteriaceae of river water origin in Turkey.