Group A streptococci persist in the tissues of the upper respiratory tract for several months after infection (1), although the number of streptococci that can be isolated from the respiratory tract usually decreases after the acute illness. The present study was designed to test the infectivity of group A streptococci isolated from the human oropharynx during the acute streptococcal infection and during the carrier state. Infectivity was determined by intranasal inoculation of monkeys, since in these animals streptococci will grow in the tissues of the oropharynx and induce an antibody response.
METHODSThe strains of group A streptococci employed were isolated from Patient 496 who developed exudative pharyngitis due to type 14 streptococci on March 21, 1951, in Cheyenne, Wyoming. He received no specific therapy and 2 weeks later was transferred by airplane to Florida (1). Cultures of the nose and oropharynx were obtained twice weekly by methods previously described (1). Colonies of bacteria showing p-hemolysis were isolated from the original sheep blood-agar plates and transferred to a second blood-agar plate. After incubation several colonies were inoculated into a blood-broth culture which was employed as the source of organisms for serological identification by the usual methods (2). A small quantity of the blood-broth culture was added to sterile milk and stored at -200 C.For inoculation of animals, 0. brain-heart infusion broth which contained one drop of sheep blood. After incubation at 370 C for 8 hours, 1 ml was transferred into a flask containing 25 ml of brainheart infusion and 0.5 ml of sheep blood. The culture was incubated at 370 C for 12 hours, chilled, and centrifuged at 2,500 rpm for 30 minutes. The sedimented organisms were resuspended in 10 ml of sterile brainheart infusion broth. With a syringe and a blunt needle, 0.5 ml of this suspension was inoculated into each nostril of unanesthetized monkeys; the procedure was repeated 24 hours later. It was demonstrated by pour-plate technique that the total inocula given to each monkey contained 400 to 780 million viable streptococci or clumps.Cultures of the oropharynx, after the second inoculation, were obtained three times weekly for 5 weeks and then once a week for 4 weeks. Cultures were streaked on sheep blood-agar plates and the number of 8-hemolytic colonies recorded. During the first week four colonies, and from subsequent cultures one colony, were selected for serological identification (2). Blood specimens for antistreptolysin 0 determinations (3) were drawn before and 3 weeks after inoculation.Eleven young male and 9 young female rhesus monkeys were housed in individual cages. Frequent oropharyngeal cultures showed that 5 animals harbored group A streptococci and 4 carried group C or G organisms. All group A, C, and G streptococci were eliminated by penicillin therapy several weeks prior to inoculation.
RESULTS