The overuse of antibiotics for the remedy of humans and animals causes antimicrobial residues released into water surfaces and wastewater. Wastewater treatment plants have been used to remove chemical and biological pollutants; however, this environment can be considered a suitable medium for the spread of resistant genes via horizontal transfer. Thus, twenty water samples were collected from the water surface of different places in the Tigris River in March 2022 in Baghdad city. Samples were analyzed to detect the resistance of  Escherichia coli against vancomycin antibiotics. Based on phenotype detection, among 25 E. coli isolates, 15 isolates were resistant to vancomycin antibiotics. While, PCR analysis showed that, among 15 vancomycin-resistant E.coli, only 8 isolates included the van-A gene, and one isolate recorded a van-B gene. Vancomycin resistance of Escherichia coli has been recorded in the environment associated with the presence of antibiotic residues released in various water environments due to the increased use of human and animal medicine, as well as farming and aquaculture for the sake of preventing and treating infections.