Coagulase-negative and deoxyribonuclease-negative mutants were isolated from Staphylococcus aureus BB by treatment with N-methyl-N'-nitro-N-nitrosoguanidine. Comparison of virulence (50% lethal dose) to mice of these six mutant strains and S. aureus BB was determined by both intravenous and intraperitoneal routes. The ratios of the 50% lethal dose of coagulase-negative mutants to that of the parental strain S. aureus BB ranged from 201 to 403 for intravenous infection and 30.7 to 52.7 for intraperitoneal infection. The virulence of deoxyribonuclease-negative mutants was essentially the same as that of S. aureus BB. When mice were immunized subcutaneously with live S. aureus BB or its deoxyribonuclease-negative mutants, the resulting protection against the intravenous challenge of S. aureus BB was remarkable. The ratios of the 50% lethal dose for the mice that were immunized by these strains to that for the untreated mice extended from 40.1 to 60.6 for intravenous infection and 6.61 to 11.5 for the intraperitoneal route. However, no effect against S. aureus BB challenge was shown in the mice that were immunized with coagulase-negative mutants. According to Bergey's Manual of Determinative Bacteriology (2), Staphylococcus aureus is coagulase and protein A positive; Staphylococcus epidermidis and Staphylococcus saprophyticus are both negative. Moreover, the former ferments mannitol anaerobically; the latter two do not. Among these species, S. aureus has been observed in diseased tissues and in pus obtained from human abscesses and has produced many kinds of extracellular toxins and enzymes. These include hemolysin, leucocidin, fibrinolysin, penicillinase, deoxyribonuclease (DNase), and phosphatase (14). Many papers have reported on the relationship between S. aureus isolated from clinical materials and its extracellular toxins and enzymes. Elston et al. (6) reported that 99.4% of the clinical S. aureus strains released coagulase, 98.8% of these coagulase-positive strains produced DNase, and 85.9% of these same coagulase-positive strains fermented mannitol. Forsgren (8) reported that 100% of the coagulase-positive strains produced DNase and 98.9% released protein A. Some studies have been made about the role of these toxins and enzymes and their relationship to the virulence of S. aureus in human t Journal article no. 8400 of the Michigan Agricultural Experiment Station.