This work aimed to focus on the synthesis of the nanohybrid compounds from silver nanoparticles (AgNPs) with Nickel/Aluminum layers of double hydroxide (LDHs) by the direct and indirect ion-exchange method at pH 6.5 as a light green-yellow colored nanostructure powder. The silver nitrate precursor was reduced to silver nanoparticles using starch as reducing and capping agents solutions at pH 8. The resultant silver nanoparticles are integrated on LDHs surface which is loaded on its surface for 12h. These nano compounds were characterized using X-ray Diffraction, Atomic Force Microscopy, Fourier Transform Infrared spectroscopy, and UV-Visible spectrophotometer. The XRD data and AFM images demonstrated that the prepared AgNPs, LDHs, and AgNPs with LDHs are nanomaterials with nano-spherical, and nano- rod and nanorod too, respectively. The antibacterial Performances of Ag NPsNi/Al–LDH manufactured by direct and indirect ionic exchange method and Ag NPs were determined against Staphylococcus aureus bacteria. Even at the 4th recycled used, the kill percentages of Ag NPsNi/Al–LDH – direct was still as higher as 100%. The growth behaviors of S. aureus were monitored by the optical density at 600nm (OD600). These newly designed of (Ag NPsNi/Al–LDH) layers may offer a promising antibacterial solution as alternative drugs to antibiotics for clinical and environmental applications. A qualitative assessment of the anti-biofilm formation effect was performed using a modified test tube method by measuring the optical density. The results demonstrated the efficient antibiofilm activity of Ag NPsNi/Al–LDH – with the direct method, its highest compared with other studied molecules and the antibiotics itself. The current data highlights, biogenic NPsNi/Al–LDH – with direct method could be used as an adjuvant for antibiotics and anti-biofilm formation in the treatment of bacterial infections.