Improved selective culture media for Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus

By modifying the previously described media tryptic soy-serum-bacitracin-vancomycin (TSBV) agar and tryptic soy-serum-bacitracin-vancomycin-fluoride (TSBVF) agar, two improved selective culture media were developed for isolation and enumeration of Actinobacillus actinomycetemcomitans (A medium) and Haemophilus aphrophilus (H medium) in oral specimens. Both media were supplemented with fusidic acid and spiramycin, and carbenicillin was also added to A medium. The growth yields of pure cultures of A. actinomycetemcomitans on A medium and of H. aphrophilus on H medium were comparable with those on the reference media. Compared with blood agar, the selective media inhibited these species about 10-fold or less. In addition, A and H media suppressed the growth of pure cultures of Capnocytophaga spp. and Neisseria spp., commonly found as contaminants on TSBV and TSBVF, 10(5) times or more compared with that on blood agar. In samples from diseased periodontal pockets, the recoveries of A. actinomycetemcomitans on A medium and H. aphrophilus on H medium equaled those on TSBV and TSBVF, respectively. In about 50% of the cultures on the reference media, contaminating bacteria were detected at levels higher than 10(4) CFU/ml of sample. The corresponding value for both A and H media was about 2%.