Systemic Doxycycline Administration in the Treatment of Periodontal Infections (II). Effect on Antibiotic Resistance of Subgingival Species

The purpose of this investigation was to determine the proportion and prevalence of doxycycline resistant species in subgingival plaque samples taken during and after doxycycline administration. 20 subjects with adult periodontitis were randomly assigned to test (n = 10) or control groups (n = 10). Saliva samples as well as subgingival plaque samples taken from the distal surface of 6 posterior teeth were collected at baseline. All subjects received full mouth SRP and the test group systemic doxycycline at the dosage of 100 mg/day for 14 days. Saliva samples and plaque samples from the distal surface of 2 randomly selected teeth were taken at 3, 7 and 14 days during and after antibiotic administration. Control subjects were sampled at the same time points. Samples were anaerobically dispersed and serially diluted in PRAS Ringer's solution and plated on enriched Trypticase soy blood agar plates with or without 4 microg/ml doxycycline. After 7 days of anaerobic incubation, colonies were counted on both sets of plates. Microbial growth was washed from the doxycycline-containing media and the species identified using 40 DNA probes and checkerboard DNA-DNA hybridization. Differences in proportions of resistant species between test and control groups were tested for significance at each time point using the Mann Whitney test and over time within each group using the Quade test. The mean % (+/-SEM) of isolates resistant to 4 microg/ml doxycycline in the plaque samples of the test subjects increased from 6+/-2 to 48+/-9% during doxycycline administration, decreasing to 25+/-6% 2 weeks later and 9+/-2% at 90 days. In saliva, the % of resistant isolates rose from 13+/-1% to 81+/-10% during doxycycline administration falling to 46+/-8% 2 weeks later and 22+/-5% at 90 days. The % of resistant isolates did not change significantly in plaque or saliva samples of the control subjects at the same time points. For all subject visits combined, the most prevalent resistant species were: Streptococcus anginosus, Streptococcus oralis, Streptococcus intermedius, Streptococcus sanguis, Streptococcus mitis, Veillonella parvula, Actinomyces gerencseriae, Streptococcus constellatus, Actinomyces naeslundii genospecies 2, Streptococcus gordonii, Eikenella corrodens and Actinomyces naeslundii genospecies 1. Doxycycline resistant strains of these species were detected in both plaque and saliva samples prior to therapy and in the control group. Despite the finding of increased resistance, approximately 50% of the organisms present at periodontal sites at the end of 14 days of doxycycline administration tested sensitive to the agent.