The virulence of pneumococcal capsular type 2 strain D39 and derivatives with mutations in the pneumolysin gene were examined in a mouse bacteremia model. In CBA/N-XID mice D39 is known to exhibit exponential growth in the blood until the death of the mice at 24 to 36 h. In contrast, PLN, a pneumolysin-deficient derivative of D39, reaches a plateau in growth that is maintained for several days. The growth patterns of D39 and PLN observed in CBA/N-XID mice were also observed in C3H/HeJ and C3H/HeOuJ mice, but not in 129/SvJ and C57BL/6J mice. These results demonstrate that the effect of pneumolysin on bacteremia is dependent on the genetic background of the mice. D39 derivatives with point mutations which abolish the cytotoxic or complement-activating properties of pneumolysin did not have major individual effects on virulence in CBA/N-XID and C3H/HeOuJ mice. A derivative with mutations affecting both the cytotoxic and complement-activating properties resulted in a modest, yet statistically significant, increase in survival time of i.v. challenged CBA/N-XID mice. However, the effect was less marked than that seen with PLN. These findings suggest that the virulence effects of pneumolysin in bacteremia must be due in part to properties other than hemolysis and complement fixation.