Infant rats were infected intranasally with mixtures of streptomycin-sensitive and streptomycin-resistant strains of Haemophilus influenzae type b and cultures of nasopharyngeal washings, blood, and cerebrospinal fluid were obtained. If from independent action, as opposed to cooperative interaction of intranasally inoculated organisms. The results also suggested that the meninges were invaded by the hematogenous route. Intranasal inoculation of infant rats with Haemophilus influenzae type b results in bacteremia and meningitis and has proved to be a useful experimental model of the human disease (1-3). These studies provided evidence that the pathogenesis of meningitis involved three sequential events: nasopharyngeal colonization, bacteremia, and central nervous system invasion, although direct invasion of the meninges by contiguous spread from the nasopharynx was not precluded. The studies also revealed that an inoculum of at least 1000 organisms was a prerequisite for the induction of bacteremia and meningitis. This observation might be explained as follows. First, it is possible that every organism in the initial inoculum behaves in a manner that is uninfluenced by the other bacteria present (independent action) (4-6). Each organism has a low probability of successfully overcoming the host defenses; however, the probability is not zero. As the dose of organisms is increased, it becomes progressively more likely that one of them will establish a local or distant focus of infection. Under these circumstances, the bacteria recovered from blood or cerebrospinal fluid (CSF), for example, would be descended from a few organisms, perhaps even a single organism of the original inoculum (4-7). Alternatively, organisms en masse may possess properties that are denied to single bacteria (cooperative action). When the inoculum attains a critical magnitude, infection ensues (8, 9). Under these circumstances, bacteria cultured from the blood or CSF would be descendents of many different organisms of the original inoculum.The following study was designed to test the theory of independent action compared to that of cooperative action in the pathogenesis of H. influenzae type b, bacteremia, and meningitis. The results also provided data relevant to the relationship between nasopharyngeal colonization, bacteremia, and invasion of the central nervous system.
MATERIALS AND METHODSThe Eagan strain of H. influenzae type b and its one-step streptomycin-resistant mutant, the media used for its growth, the methods of intranasal inoculation, the technique of blood and CSF sampling, and the quantitation of bacteria in blood and CSF were identical to those used previously (1-3). Natural litters of 5-day-old rats (Sprague-Dawley, strain COBS/CD) were obtained from Charles River Laboratories, Wilmington, MA. Inocula containing an approximately equal number of the streptomycin-sensitive (SMS) and streptomycin-resistant (SMR) variants were prepared as follows. Each variant was grown overnight on solid medium at 370. A few colonies ...