Bovine Respiratory Disease (BRD) is a worldwide health problem and an important cause of disease and death in feedlot cattle. BRD is caused by several pathogens, including bovine herpesvirus-1 (BHV-1), which is the most prevalent virus associated with BRD. Recently, infection with highly pathogenic avian influenza (HPAI) H5N1 clade 2.3.4.4b viruses in dairy cattle have also been reported. Currently, treatment options against either virus in cattle remains suboptimal. We are currently investigating host-targeted therapies for treatment of BHV-1 and/or IAV infections in cattle. RIG-I is an intracellular pattern recognition receptor and synthetic RIG-I agonists have been previously shown to upregulate interferon stimulated genes (ISGs) and protect against viral infections in human cells in vitro. We demonstrate that pre-treatment of bovine MDBK cells with RIG-I agonists also results in potent inhibition of IAV and BHV-1. Previous studies have shown that human and mouse myxovirus resistance protein (Mx)1and/or Mx2 are key ISGs induced by RIG-I agonists and associated with its antiviral activity. Given that very little is known regarding induction and antiviral activity of Mx in cattle, we first confirmed that bMx1 and bMx2 were rapidly induced in MDBK cells in response to bovine interferon (IFN)-a. Second, we demonstrated that MDBK cells with inducible overexpression of bMx1, but not bMx2, inhibited growth of several influenza A viruses (IAV) of different subtypes, including an avian HPAI H5N1 virus from the clade 2.3.4.4b. Finally, we show that mutation of key residues in bMx1 altered their distribution and/or localisation in bovine cells and that some mutations also impacted anti-IAV activity. Current studies are investigating if bMx1/bMx2 also mediate antiviral activity against BHV-1. These studies highlight the importance of studying antiviral immunity against bovine viruses, including those with zoonotic potential, as well as the potential for development of host-directed antivirals with broad antiviral activity in cattle.