A Selective Medium for <i>Actinobacillus actinomycetemcomitans</i> and the Incidence of the Organism in Juvenile Periodontitis

A selective medium, malachite green bacitracin agar, was developed for the isolation of Actinobacillus actinomycetemcomitans from subgingival plaque of periodontally diseased patients. The medium consisted of Trypticase soy agar 40 gm/liter, bacitracin 128 micrograms/ml, malachite green 8 micrograms/ml and 5% defibrinated sheep blood. The medium, when incubated in an atmosphere of air plus 10% CO2 for 5 days, permitted greater than 80% recovery of pure cultures of A. actinomycetemcomitans when compared with a nonselective medium. The most frequent contaminant in plaque samples from different clinical conditions was Haemophilus aphrophilus. Decomposition of H2O2 was useful in differentiating these two species. Clinical studies employing the malachite green bacitracin medium revealed a significant association between the presence of the organism, A. actinomycetemcomitans and juvenile periodontitis.