Laser ablation from a solid target assists the synthesis of nanoparticles (NPs) as an alternative physical process. This work is attempt to investigate the morphological properties of silver nanoparticles (AgNPs) synthesized by femtosecond pulsed laser ablation in different liquid medium Double Distilled Water (DDW), Deionized Water (DIW), Tetrahydrofuran (THF), and Dimethylformamide (DMF). XRD diffraction peaks that corresponds to (111), (200), (220) and (311) planes confirm the synthesized of AgNPs. The absorption spectra confirm the existence surface plasmon resonance (SPR) peak of AgNPs. The direct optical energy band gap (Eg) values have been calculated according to Taucs equation and equal to 3.97, 3.82, 3.86, and 3.88 eV for synthesized AgNPs by Fs PLAL at different medium DDW, DIW, THF, and DMF, respectively. Also, the influence of different liquid medium on the antibacterial efficiency was studied. In vitro Antibacterial activity of synthesized AgNPs was carried out against four types of bacteria employing agar plate technique. The experiments demonstrated that ablation efficiency, stability, and antibacterial efficiency of AgNPs in DIW medium are higher than those prepared in DDW, THF, and DMF medium.