Leather is often used to make comfortable shoes due to its soft and breathable nature. However, its innate ability to retain moisture, oxygen and nutrients renders it a suitable medium for the adsorption, growth, and survival of potentially pathogenic microorganisms. Consequently, the intimate contact between the foot skin and the leather lining surface in shoes, which are subject to prolonged periods of sweating, may result in the transmission of pathogenic microorganisms and cause discomfort for the wearer. To address such issues, we modified pig leather with silver nanoparticles (AgPBL) that were bio-synthesized fromPiper betleL. leaf extract as an antimicrobial agent via the padding method. The evidence of AgPBL embedded into the leather matrix, leather surface morphology and element profile of AgPBL-modified leather samples (pLeAg) was investigated using colorimetry, SEM, EDX, AAS and FTIR analyses. The colorimetric data confirmed that the pLeAg samples changed to a more brown color with higher wet pickup and AgPBL concentration, owing to the higher quantity of AgPBL uptake onto the leather surfaces. The antibacterial and antifungal activities of the pLeAg samples were both qualitatively and quantitatively evaluated using AATCC TM90, AATCC TM30 and ISO 16187:2013 test methods, approving a good synergistic antimicrobial efficiency of the modified leather againstEscherichia coliandStaphylococcus aureusbacteria, a yeastCandida albicansand a moldAspergillus niger. Additionally, the antimicrobial treatments of pig leather did not negatively impact its physico-mechanical properties, including tear strength, abrasion resistance, flex resistance, water vapour permeability and absorption, water absorption and desorption. These findings affirmed that the AgPBL-modified leather met all the requirements of upper lining according to the standard ISO 20882:2007 for making hygienic shoes.