Staphylococcus aureus ( S. aureus) is often associated with cases of bovine mastitis, a common disease in dairy herds that causes economic losses to the farmer and industry. In addition, S. aureus is often associated with outbreaks of food poisoning, as some strains can produce staphylococcal enterotoxins (SEs). SEs are a major cause of gastroenteritis resulting from the consumption of contaminated foods, including milk and dairy products. Although S. aureus is destroyed by the heat treatment of milk pasteurization, its enterotoxins are not, remaining active in processed products. One way to control this bacterium in cases of mastitis is the use of antibiotics, which is a concern for public health and one of the biggest challenges for the food industry, as antibiotic residues can be present in milk and meat. A possible alternative for the biological control of this pathogen is the use of lytic bacteriophages or their endolysins. Thus, this proposal aims to evaluate the use of lytic bacteriophage endolysins to control S. aureus in the milk production chain. Initially, the endolysin gene from two bacteriophages, that were previously isolated by our group in Embrapa Dairy Cattle, was cloned. The two bacteriophages showed the highest lytic capacity among the six bacteriophages isolated and tested against 100 strains of S. aureus. The recombinant proteins are being purified, and will subsequently be characterized and evaluated in comparison to a mixture of bacteriophages regarding their teat surface disinfectant potential, aiming at controlling the bovine staphylococcal mastitis, the hand hygiene of handlers and the development of S. aureus in Minas cheese. It is expected at the end of the work to have knowledge of the efficacy of a mixture of bacteriophages and a recombinant protein and to be able to recommend the best of them as an antimicrobial alternative to chemicals in controlling S. aureus in the dairy chain with benefits to livestock farmers, industry and consumers.