The current study characterizes the genetic distribution of virulence and antimicrobial resistance of Streptococcus lutetiensis and Streptococcus equinus isolated from cows with clinical mastitis using whole genome sequencing (WGS). Although they are not the protagonist species within the genus Streptococcus, recent studies have isolated these species associated with bovine mastitis. In addition, these species are reported and isolated from humans and other animals. A total of four strains of S. lutetiensis and one of S. equinus were isolated from five cows with identified cases of clinical mastitis at a dairy farm near Ithaca, New York. Nineteen genes associated with antimicrobial resistance and 20 genes associated with virulence were identified in the analyzed strains. All strains presented genes associated with resistance: alr, ddl, gdpD, kasA, murA, lsa(E), msr(D), mef(A), gidB, and LiaF. Resistance genes associated with several different classes of antibiotics have also been reported. Sixteen virulence-associated genes were identified in all strains. Based on our findings, we conclude that the studied species have the potential to cause mastitis in cattle, and further studies are important to elucidate their role.