Despite the presence of modern antibacterial drugs, bacterial infections are still a major threatening problem due to the enormous increase in multi-drug-resistant bacteria. Nanoparticles have been extensively used as an applicable and safe alternative to antibiotics. The present study aimed to explore the inhibitory effect of silver nanoparticles on Extended Spectrum Beta lactamase (ESBL) producing E. coli and Klebsiella spp. in vitro as well as their effect on the expression of antibiotic resistance genes. Different samples (i.e., wound swabs, Fecal swabs, and urine samples) were collected from dogs and cats. Phenotypic and molecular identification, antibiotic susceptibility testing, and double-disk synergy test were carried out for the identification of ESBL producing E. coli and Klebsiella spp. Silver nanoparticles were tested for their in vitro antibacterial potential and there were reports of their minimum inhibitory concentration and minimum bactericidal concentration. Moreover, the effect of silver nanoparticles on the expression of antibiotic resistance genes (i.e., blaTEM, blaSHV, and blaCTX) was assessed as well as their effect on the structural integrity of the bacterial cells using Scanning Electron Microscope (SEM). Results revealed that 23 isolates (19.16%) (E. coli=17, Klebsiella spp.=6) were confirmed as ESBL producing. Silver nanoparticles indicated a promising antibacterial effect where the minimum inhibitory concentration of AgNPs for ESBL producing E. coli was measured as 0.31 mg/ml, and 0.62 mg/ml for ESBL-producing Klebsiella spp., while the minimum bactericidal concentration of ESBL-producing E. coli and Klebsiella spp. was reported as 0.15 mg/ml and 0.3 mg/ml, respectively. Consequently, the expression of antibiotic resistance genes was downregulated in both bacteria species and there was a noticeable toxic effect of AgNPs on E. coli and Klebsiella spp. cells which was investigated using SEM. It can be concluded that silver nanoparticles have a promising antibacterial activity and could be considered an applicable alternative for the control of ESBL producing bacteria.