Article

Selective Medium for Isolation of Actinobacillus actinomycetemcomitans

Journal of Clinical Microbiology (Impact Factor: 3.99). 05/1982; 15(4):606-9.
Source: PubMed

ABSTRACT

A selective medium, TSBV (tryptic soy-serum-bacitracin-vancomycin) agar, was developed for the isolation of Actinobacillus actinomycetemcomitans, TSBV agar contained (per liter) 40 g of tryptic soy agar, 1 g of yeast extract, 100 ml of horse serum. 75 mg of bacitracin, and 5 mg of vancomycin. The TSBV medium suppressed most oral species and permitted significantly higher recovery of A. actinomycetemcomitans than nonselective blood agar medium. The distinct colonial morphology and positive catalase reaction of A. actinomycetemcomitans easily distinguished this bacterium from Haemophilus aphrophilus, Capnocytophaga species, and a few other contaminating organisms. With the TSBV medium, even modestly equipped laboratories will be able to isolate and identify A. actinomycetemcomitans from clinical specimens.

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    • "4-Brucella agar (BA) plates enriched with 5% defibrinated sheep blood and 5 mg/ml clindamycin to assess Eikenella corrodens. 5-TSBV (trypticase soy, serum, bacitracin 75 mg/ml, vancomycin 5 mg/ml) as a selective media for Aggregatibacter acti- nomycetemcomitans[26]the plates were incubated at 37 C microaerophilically. All isolates were subcultured to obtain purified cultures then identified by API-ZIM System supplied by (BioM erieux,France). "
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    • "In the laboratory, samples were vortexed and seeded on blood agar (horse blood) and Tryptic Soy Serum Bacitracin Vancomycin agar (TSBV) (horse serum) [19] medium for identification of A. actinomycetemcomitans and other bacteria with known in-or low sensitivity to MET [15]. The seeded samples were cultured at 35 C, aerobically, as well as in 5% CO 2 in air, for 5 days, and bacterial colonies described and identified by physical appearance and biochemical tests [20]. "
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    • "These antimicrobial concentrations represent non-susceptible/resistant breakpoint concentrations against anaerobic bacteria for amoxicillin, metronidazole, and clindamycin as recommended by the Clinical and Laboratory Standards Institute (CLSI) (Clinical & Laboratory Standards Institute 2012b), and for doxycycline disk diffusion testing as recommended by the French Society for Microbiology (Comit e de l'Antibiogramme de la Soci et e Franc ßaise de Microbiologie 2010). Direct colony suspensions (equivalent to a 0.5 McFarland standard ) of pure A. actinomycetemcomitans isolates from selective TSBV plates were subcultured onto these media as their recognition is frequently obscured within mixed bacterial populations (Slots 1982). In vitro resistance to the antibiotic breakpoint concentrations of doxycycline, amoxicillin, metronidazole , or clindamycin was recorded when test species growth was noted on the respective antibiotic-supplemented EBBA plates (Slots et al. 1988; Feres et al. 1999; van Winkelhoff et al. 2000b; Rams et al. 2011b, 2012). "
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