Selective Medium for Isolation of Actinobacillus actinomycetemcomitans

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


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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    • "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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    ABSTRACT: Objective: The benefit of full-mouth disinfection (FDIS) over traditional scaling and root planing (SRP) in the treatment of chronic, destructive periodontitis remains equivocal and it is not known whether the use of adjunctive antibiotics may enhance the effect of FDIS. Therefore, the aim of this study was to evaluate the effect of conventional SRP completed over 21 days or 1-day FDIS, with or without systemically delivered adjunctive metronidazole (MET) on the presence of P. gingivalis and T. forsythia after 3 and 12 months. Materials and methods: One hundred and eighty-four patients with moderate-to-severe periodontitis were randomly allocated to one of four treatment groups; (1) FDIS+MET; (2) FDIS+placebo; (3) SRP+MET; (4) SRP+placebo. Prior to treatment, pooled subgingival samples were obtained from the five deepest pockets. The same sites were sampled again 3 and 12 months after treatment. All samples were analyzed for P. gingivalis and T. forsythia by PCR, whereas A. actinomycetemcomitans and other bacteria were identified by culture techniques. Results: At baseline, 47% of the samples were positive for P. gingivalis, while almost all samples were positive for T. forsythia. The occurrence of P. gingivalis and T. forsythia was significantly reduced at 3 and 12 months after treatment in the FDIS+MET group, but not in the other treatment groups. Conclusion: FDIS+MET had a significant effect in patients with P. gingivalis and T. forsythia, resulting in a significant reduction in number of patients where these micro-organisms could be detected at 3 and 12 months post-therapy.
    Acta Odontologica Scandinavica 01/2015; 73(4):1-9. DOI:10.3109/00016357.2014.920106 · 1.03 Impact Factor
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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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    ABSTRACT: Objectives: Because antimicrobial therapy is often employed in the treatment of infectious dental implant complications, this study determined the occurrence of in vitro antibiotic resistance among putative peri-implantitis bacterial pathogens. Methods: Submucosal biofilm specimens were cultured from 160 dental implants with peri-implantitis in 120 adults, with isolated putative pathogens identified to species level, and tested in vitro for susceptibility to 4 mg/l of doxycycline, 8 mg/l of amoxicillin, 16 mg/l of metronidazole, and 4 mg/l of clindamycin. Findings for amoxicillin and metronidazole were combined post-hoc to identify peri-implantitis species resistant to both antibiotics. Gram-negative enteric rods/pseudomonads were subjected to ciprofloxacin disk diffusion testing. Results: One or more cultivable submucosal bacterial pathogens, most often Prevotella intermedia/nigrescens or Streptococcus constellatus, were resistant in vitro to clindamycin, amoxicillin, doxycycline, or metronidazole in 46.7%, 39.2%, 25%, and 21.7% of the peri-implantitis subjects, respectively. Only 6.7% subjects revealed submucosal test species resistant in vitro to both amoxicillin and metronidazole, which were either S. constellatus (one subject) or ciprofloxacin-susceptible strains of gram-negative enteric rods/pseudomonads (seven subjects). Overall, 71.7% of the 120 peri-implantitis subjects exhibited submucosal bacterial pathogens resistant in vitro to one or more of the tested antibiotics. Conclusions: Peri-implantitis patients frequently yielded submucosal bacterial pathogens resistant in vitro to individual therapeutic concentrations of clindamycin, amoxicillin, doxycycline, or metronidazole, but only rarely to both amoxicillin and metronidazole. Due to the wide variation in observed drug resistance patterns, antibiotic susceptibility testing of cultivable submucosal bacterial pathogens may aid in the selection of antimicrobial therapy for peri-implantitis patients.
    Clinical Oral Implants Research 04/2013; 25(1). DOI:10.1111/clr.12160 · 3.89 Impact Factor
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    • "de Man-Rogosa- Sharpe (MRS) agar was used for the isolation of Lactobacilli [14]. Tryptone soya agar which was supplemented with yeast extract (0.1%), horse serum (10%), bacitracin (75µg/ml) and vancomycin (5µg/ml) (TSBV), was used for the isolation of A. actinomycetemcomitans [15]. The plates were incubated in a microaerobic atmosphere (10% CO 2 ) for 72 hours at 37 0 C. "
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    ABSTRACT: Introduction: The purpose of the present study was to evaluate the effect of a probiotic (Inersan®) alone, a combination of the probiotic with doxycycline and doxycycline alone on aggressive periodontitis patients. Methods: Thirty patients who satisfied the inclusion and exclusion criteria, were assigned to one of the above mentioned three groups by using block randomization. The clinical and the microbiological parameters were recorded on day 0, at 2 weeks and at 2 months. On day 0, before recording the clinical parameters, 0.5 ml of unstimulated saliva was collected for the evaluation of the microbiological parameters. The clinical parameters which were recorded were the plaque index, the gingival index, the probing pocket depth and the clinical attachment level. The microbiological parameters which were recorded were Lactobacilli and Aggregatibacter actinomycetemcomitans. After this, Scaling and Root Planing (SRP) was performed on day 0. Two weeks after the SRP, the patients were recalled for the saliva sample collection and for the evaluation of the clinical parameters. On the same day, medications were given to the patients to be taken for fourteen days according to the group which they belonged to (Group A - probiotic alone, Group B - a combination of the probiotic with doxycycline, Group C - doxycycline alone). The patients were then recalled at two months for the saliva sample collection and for the evaluation of the clinical parameters. Results: The administration of the probiotic alone, a combination of the probiotic with doxycycline and doxycycline alone, resulted in a decrease in the plaque index, the gingival index, the probing pocket depth and the clinical attachment level at 2 months, which was statistically significant (p < 0.05). The A. actinomycetemcomitans count tended to decrease in all the three groups at 2 months, which was statistically non-significant (p > 0.05). The Lactobacilli count tended to increase significantly in the probiotic alone group (p < 0.05). Conclusion: Probiotics have a future in the treatment of aggressive periodontitis, as antibiotics are prescribed most of the time. These antibiotics can lead to the emergence of drug resistant micro-organisms and they can also disturb the beneficial microflora of the body. Thus, as an alternative to antibiotics, probiotics can be used, as they repopulate the beneficial microflora and reduce the pathogenic bacteria.
    03/2013; 7(3):595-600.
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