Streptococcus pneumoniae is a causative agent for bacterial pneumonia, otitis media, meningitis, and bacteremia. Mechanisms for acquisition of iron by this organism under low-iron conditions were investigated. Siderophore production was not detected by either chemical or biological methods. Its utilization of iron-containing compounds found in human hosts was tested. Both hemin and hemoglobin supported the full growth of S. pneumoniae in a culture lacking other iron sources, while lactoferrin and transferrin failed to do so. A mutant defective in hemin utilization was isolated and was less virulent than wild-type S. pneumoniae in experimental animals.