Silver nanoparticles (Ag-NPs) have unique properties as antibacterial effects against locally isolated clinical Escherichia coli.  In this study, the evaluated the antibacterial activity of AgNPs, which were synthesized by laser ablation, against locally isolated clinical  Escherichia coli on nutrient agar media in vitro. Then assessed the toxicity of the bactericidal dose in albino rats in vivo with hematological, liver, and kidney functions as vital parameters. AgNPs were synthesized by pulsed laser ablation in liquid (PLAL).  AgNPs’ shape and nano size were characterized by atomic force microscopy (AFM), UV-vis spectroscopy, and scanning electron microscopy (SEM). UV peak absorption was observed at 434 nm, which produced an average particle diameter of 28.09 nm. The SEM and AFM topography images of the synthesized nanoparticles showed that they had a spherical shape. Also studied was the toxicity of the synthesized AgNPs in vivo. Four groups of albino rats were used. The liver and kidneys were found to be the main organs that accumulate AgNP. In this study, we examined the influence of intraperitoneally injected AgNPs on blood parameters (complete blood picture) and liver and kidney functions of albino rats. Blood ALP and serum test (GOT and GPT), serum urea and serum creatinine were analyzed at 15, 30, and 60 days after injection of 200 mg/kg of PLAL-synthesized AgNP. The results did not indicate statistically significant differences between the control and treatment groups in blood picture and enzyme levels tested that indicate proper liver function. Additionally, no differences in kidney functions were found during varying time intervals in the blood of rats. Thus, PLAL-synthesized AgNPs could inhibit Escherichia coli bacterial growth.