Bovine mastitis is a multifactorial disease that produces important losses in livestock and dairy industry. One of the main factors that causes this disease are microorganisms such as Staphylococcus aureus, which has multiresistance to antibiotics and virulence factors such as its ability to internalize into bovine mammary epithelial cells (bMECs), which allows bacteria to evade the host's immune system and persist inside the cell. However, coagulasenegative Staphylococci (CNS) have a significant role as bovine mastitis pathogens. S. agnetis is an emergent CNS pathogen isolated from bovine mastitis, whose capacity to internalize into bMECs is unknown. In the present work we characterized an isolate of S. agnetis from bovine mastitis, which was resistant to penicillin, dicloxacillin and ampicillin; also, the isolate showed resistance to methicillin through a mechanism that may be related to the presence of the mecA gene. In addition, S. agnetis internalized into bMECs (10% relative to S. aureus (ATCC 27543), which could be related to the presence of the fnbpA, clfB and spa genes. This is the first study that demonstrates the ability of internalization of S. agnetis into bMECs, which is a virulence factor that may represent a major problem in the management of bovine mastitis.