Prevalence of ß-lactamase-producing bacteria in human periodontitis.

BACKGROUND AND OBJECTIVE: Beta-lactam antibiotics prescribed in periodontal therapy are vulnerable to degradation by bacterial ß-lactamases. This study evaluated the occurrence of ß-lactamase-positive subgingival bacteria in chronic periodontitis subjects of USA origin, and assessed their in vitro resistance to metronidazole at a breakpoint concentration of 4 µg/mL. MATERIAL AND METHODS: Subgingival plaque specimens from deep periodontal pockets with bleeding on probing were removed from 564 adults with severe chronic periodontitis before treatment. The samples were transported in VMGA III and then plated onto: (i) nonselective enriched Brucella blood agar (EBBA) and incubated anaerobically for 7 d; and (ii) selective trypticase soy-bacitracin-vancomycin (TSBV) and incubated for 3 d in air + 5% CO2 . At the end of the incubation periods, the bacterial test species were identified and quantified. Specimen dilutions were also plated onto EBBA plates supplemented with 2 µg/mL of amoxicillin, a combination of 2 µg/mL of amoxicillin plus 2 µg/mL of the ß-lactamase inhibitor clavulanic acid, or 4 µg/mL of metronidazole, followed by anaerobic incubation for 7 d. Bacterial test species presumptively positive for ß-lactamase production were identified by growth on EBBA primary isolation plates supplemented with amoxicillin alone and no growth on EBBA primary isolation plates containing both amoxicillin plus clavulanic acid. A subset of such isolates was subjected to nitrocefin-based chromogenic disk testing to confirm the presence of ß-lactamase activity. In vitro resistance to 4 µg/mL of metronidazole was noted when growth of test species occurred on metronidazole-supplemented EBBA culture plates. RESULTS: Two-hundred and ninety-four (52.1%) of the study subjects yielded ß-lactamase-producing subgingival bacterial test species, with Prevotella intermedia/nigrescens, Fusobacterium nucleatum and other Prevotella species most frequently identified as ß-lactamase-producing organisms. Of the ß-lactamase-producing bacterial test species strains recovered, 98.9% were susceptible in vitro to metronidazole at 4 µg/mL. CONCLUSION: The occurrence of ß-lactamase-positive subgingival bacterial species in more than half of the subjects with severe chronic periodontitis raises questions about the therapeutic potential of single-drug regimens with ß-lactam antibiotics in periodontal therapy. The in vitro effectiveness of metronidazole against nearly all recovered ß-lactamase-producing subgingival bacterial species further supports clinical periodontitis treatment strategies involving the combination of systemic amoxicillin plus metronidazole.

Peptostreptococcus micros in human periodontitis.

Peptostreptococcus micros is a recognized pathogen in medical infections, and its association with progressive periodontitis was examined in this study. P. micros was isolated from paper-point subgingival samples on anaerobic enriched blood agar plates and identified on the basis of cellular and colonial morphology and selected biochemical tests. In a cross-sectional study involving 907 people with advanced adult periodontitis, 127 with early-onset periodontitis, and 12 with localized juvenile periodontitis, P. micros in these patient groups occurred with a prevalence of 58-63%. In culture-positive patients, P. micros averaged 12-15% of total viable counts. P. micros demonstrated similar occurrence and proportional recovery in all age groups. In a longitudinal study of 91 adult periodontitis patients on maintenance therapy, P. micros demonstrated a significantly higher prevalence in disease-active than in disease-inactive patients (47% vs 14%). Mechanical subgingival debridement and 0.12% chlorhexidine pocket irrigation was unable to eradicate subgingival P. micros from 18 of 22 adult periodontitis patients. In vitro antimicrobial susceptibility testing showed P. micros to be sensitive to therapeutic levels of penicillin, clindamycin and metronidazole. Our findings indicate that P. micros is a potential pathogen in adult periodontitis. The methods for its eradication from subgingival sites remain to be determined.

In vitro antimicrobial sensitivity of enteric rods and pseudomonads from advanced adult periodontitis.

The prevalence and in vitro antimicrobial sensitivity of isolates of enteric rods and pseudomonads was examined in 844 adult periodontitis patients. These organisms were recovered from 13.5% of the study subjects. Ciprofloxacin exhibited the highest inhibitory activity of the 14 oral antimicrobial agents tested. Beta-lactam antibiotics were largely ineffective, and tetracycline failed to inhibit most Pseudomonas species. In vitro sensitivity data suggest that a subgingival microbiota comprising mainly streptococci would result from therapy that combined ciprofloxacin and metronidazole. Since streptococci may inhibit the growth of several putative periodontal pathogens, populations of "beneficial" streptococci in the periodontal pocket might constitute a very attractive therapeutic outcome. Controlled clinical studies are needed to clarify the possible role of ciprofloxacin in the treatment of destructive periodontitis.