Selective medium for the isolation of Haemophilus aphrophilus from the human periodontium and other oral sites and the low proportion of the organism in the oral flora

Abstract

We developed a medium for the selective recovery of Haemophilus aphrophilus. The medium, designated TSBVF, was composed of 4% tryptic soy agar, 10% heat-inactivated horse serum, 75 micrograms of bacitracin per ml, 5 micrograms of vancomycin per ml, and 50 micrograms of sodium fluoride per ml. TSBVF yielded a threefold higher recovery of oral H. aphrophilus than did chocolate agar with 75 micrograms of bacitracin per ml, which is a medium routinely used to diagnose human Haemophilus infections. H. aphrophilus and the few contaminating organisms on TSBVF were readily distinguished on the basis of colony morphology. The H. aphrophilus isolates exhibited variable fermentation of raffinose and dextrin but otherwise were biochemically similar. In a clinical study, H. aphrophilus was frequently recovered from supragingival plaque and saliva and occasionally from buccal mucosa and the tonsils. It was also isolated from 29 of 56 subgingival sites in 11 of 14 subjects. Its proportion of the subgingival microflora averaged 0.13% for healthy periodontal sites, 0.05% for adult periodontitis lesions, and 0.03% for localized juvenile periodontitis lesions. We concluded that H. aphrophilus is an indigenous bacterium of the human oral cavity. It occurs in low proportions in subgingival plaque and plays no apparent role in advanced periodontal disease in humans.