The existence and prevalence of multi-drug resistant extended-spectrum βlactamase (ESBL)-producing bacteria in the river water is a major cause of numerous diseases worldwide. In this study, the molecular characterization of antimicrobial-resistant bacteria producing ESBL encoding genes was investigated for a better understanding of the risk factors and public health issues. The potential ESBL-producing bacterial species were detected using 16S ribose ribonucleic acid (rRNA) polymerase chain reaction (PCR). This served as a screening step to detect potential ESBLs encoding genes which were confirmed by phenotypes (DDST and E-test) and genotypes (PCR) assays with the presence of the bla genes; TEM, CTX-M, OXA-1 and SHV. Furthermore, all the confirmed bacterial isolates producing ESBL encoding genes were analysed for antibiotic susceptibility patterns against 10 different classes of antibiotics as a choice of therapy using antibiotic susceptibility testing (AST) by the disc diffusion method. The highest bacterial isolates were confirmed as Enterobacteriaceae (89.5%), predominantly Escherichia coli and Klebsiella pneumoniae. Among the 20 bacterial isolates, 12 (60 %) bacteria possessed one or more ESBLs encoding genes. Relatively high occurrence rates of β-lactamase genes; bla TEM 35%, bla SHV 20%, bla OXA-15% and bla CTX-M 10% were recorded. All the ESBLs encoding isolates showed high resistance to penicillin’s, third-generation cephalosporins, monobactams, cephamycins and carbapenems. High occurrences of ESBLs producing bacteria in the environment pose a threat to exposed communities. Therefore, early detection of MDR beta-lactamase mediated resistance genes are essential to avoid numerous diseases due to the dissemination of ESBLs producing strains.