The use of antibiotics in food-producing animals for infections treatment, metafilaxis and, although not allowed in Europe, as growth enhancer is responsible for the presence of antibiotic residues in animal derived foodstuffs. For this reason, it is very important to perform a monitoring. Muscle samples from bovine, pig, poultry, turkey and fish, as well as bovine milk and hen's egg samples, deriving from 444 farms of both Umbria and Marche regions (Italy) were analyzed by well-established and validated analytical methods in order to evaluate the presence or not of antibiotic residues (penicillins, quinolones, tetracycline and sulphonamides). The samples were collected during 2012-2021 period of time. In total, 15/2,354 samples resulted positive to the analyses. The amount of antibiotics found in the 15 samples resulted below the maximum residue limit fixed by EU Regulation 37/2010 and for this reason considered compliant. Despite irregular samples were not found, the presence of antibiotic residues in foodstuff represents a risk for public health as they are responsible for the selection of resistant strains contributing to antimicrobial resistance problem spread. In the present work, this aspect was evaluated in relation to the results obtained from the analyzed samples coming from Umbria and Marche regions.