A coagulase-negative variant of Staphylococcus aureus was identified in a herd of 250 lactating dairy cows. During testing of the entire herd, this strain of S aureus was isolated from aseptically collected milk samples of 25 cows. Cows with intramammary infections attributable to coagulase-negative S aureus had an increased somatic cell count in their milk, which was indicative of mastitis infection. Speciation of the Staphylococcus organisms was made, using a series of biochemical tests. A strain of a coagulase-positive S aureus also caused intramammary infections in the herd and shared identical biochemical characteristics with the coagulase-negative strain. Moreover, both strains could not be typed by the use of the International Set of Bovine Phages. Analysis of these findings indicated that a coagulase-negative variant of S aureus can cause intramammary infections in cattle, coagulase-negative variants of S aureus that cause mastitis can be more prevalent in herds than coagulase-positive variants, and clinicians should avoid misclassifying coagulase-negative S aureus as organisms that are clinically unimportant.