In modern medical processes, new antimicrobial drugs are being widely used in order to rapid and essential eradication of resistant bacterial strains. Broad-spectrum beta-lactamases are enzymes originated from mutations in the common plasmid genes mediators for beta-lactamases such as TEM-1, TEM-2, and SHV-1 are transferred among bacterial species. The genes coding the production of ESBL are often connected to other resistant genes causing resistance to drugs with broad spectrums. Therefore, there are different experimental techniques to detect ESBLs.This study aimed to investigate the presence of blaSHV and bla TEM genes coding beta-lactamase enzymes inEscherichia coli isolated from Zarjob River in Gilan Province, their resistance to different antibiotics, and their stereotyping. In order to isolate and identifyE. coli, 25 water samples were taken from Zarjob River and analyzed by MPN test and biochemical tests. Antibiotic sensitivity testing with Kirby-Bauer method, i.e. disk diffusion on 25E. coli isolates, and also serotyping with agglutination on a slide were performed. To detectblaSHV and blaTEM genes in E. coli, PCR technique was used and all results were analyzed by using the software SPSS. Among the 25 E. coli isolates, 18 cases (72%) had the gene blaTEM and 15 cases (60%) had the geneblaSHV. The results of polyvalent antiserums serotyping were observed as Group I (O26, O55, O111), Group II (O88, O127), Group III (O44, O125, O128), and Group IV (O20, O114).