Highly pathogenic avian influenza H5N1 viruses have circulated globally since 1996. In recent years, clade 2.3.4.4b H5N1 viruses have crossed the species barrier into multiple species, including dairy cattle. Previous studies have demonstrated that Holstein cows are susceptible to H5N1 infection. To investigate whether Jersey cows are susceptible to H5N1 infection, we inoculated three Jersey lactating cows intranasally and intramammarily with a human H5N1 influenza virus. Following inoculation, milk production rapidly decreased, and milk samples exhibited a colostrum-like appearance. Milk production remained low for at least seven days. Infection was confirmed by high levels of viral RNA detected in external and internal swabs of infected teats and milk samples. Conversely, low levels of viral RNA were detected in the respiratory tract of infected cows. These findings confirm that Jersey cows are susceptible to H5N1 infection and establish them as a valuable experimental model for studying disease pathogenesis and vaccine development.