Silver nanoparticles (AgNPs) are of interest due to their antimicrobial activity and are seen as potential candidates to replace antibiotics in animal husbandry. A few studies have focused on this new application, but they lack any considerations about residual accumulation of AgNPs in edible animal tissues and animal products. In this research, a 22 day in vivo study was carried out by oral administration of 20 nm spherical PVP coated AgNPs to hens. Six doses of approximately 1 mg kg-1 of AgNPs-PVP each were administered to animals throughout the experimentation. Atomic absorption spectroscopy (AAS) was used for quantitative determination of residual total Ag in different organs and matrices. The analyses showed that Ag accumulates in livers (concentration ranging from 141 μg kg-1 to 269 μg kg-1) and yolks (concentration ranging from 20 μg kg-1 to 49 μg kg-1) but not in muscles, kidneys, and albumen belonging to hens of the treated group (tG2). Ag was not detected in animals of the control group (uG1) (i.e., total Ag < LOD = 10 μg kg-1). Single particle inductively coupled plasma mass spectrometry (spICP-MS) and scanning electron microscopy with energy dispersive X-ray detection (SEM-EDX) were employed to elucidate the presence of AgNPs in livers and yolks belonging to tG2 animals. spICP-MS highlighted that part of residual Ag found in livers (about 5-20%) is in NP form with an average dimension of approximately 20 nm. SEM-EDX technique confirmed the presence of AgNPs only in livers of treated animals. The results show that feeding AgNPs to hens may become a source of consumer exposure to AgNPs. As far as we know this is the first study showing transfer of AgNPs or reaction products thereof from animal feed to animal products.