Observance of hygiene precautions in livestock breeding is the basis for ensuring a high level of protection of human health against zoonoses and food-borne diseases. Appropriate level of biosecurity leads to improved health and animal performance, which will reduce the use of antimicrobial agents and also increase the food safety at the beginning of the food production chain. This paper is focused on an overview of selected methods, which can be used for determination of residues of inhibitors in food and raw materials of animal origin.