The tube coagulase test, an invaluable laboratory tool for identifying Staphylococcus aureus, is most often done using rabbit plasma. However, there is evidence that depending on the origin of the isolates, other plasmas may be superior. The current study sought to compare the utility of dog and rabbit plasma in the coagulase test for S. aureus isolated from canine (n = 28), bovine (n = 29), and human (n = 30) hosts. Overall, coagulation times were significantly faster for dog (2.38 hr) than rabbit (3.19 hr) plasma. When coagulation times were compared by isolate origin, no significant differences were found for rabbit plasma, whereas bovine isolates clotted dog plasma significantly faster (1.86 hr) than canine (2.79 hr) or human (2.38 hr) isolates. Investigators should be aware that rabbit plasma may not be the ideal coagulase-testing medium for S. aureus from all sources.