Tetracycline in combination with scaling and root planing is frequently used to treat refractory periodontal disease. This study examined tetracycline resistance in bacteria recovered from periodontal pockets of patients with refractory periodontitis. Bacterial isolates resistant to 10 micrograms/ml of tetracycline were isolated from plaque samples of 17 patients, of whom 6 had received tetracycline within 8 weeks prior to sampling. Minimal inhibitory concentrations (MICs) of tetracycline and minocycline were determined by agar dilution. In the 6 patients who had received tetracycline, a mean of 22.9% (+/- 38.2) of the total cultivable subgingival flora were resistant to tetracycline, compared with a mean of 7.2% (+/- 8.5) in the untreated group. Although various organisms were isolated, in most patients, the tetracycline-resistant organisms were dominated by Streptococcus spp. Overgrowth of Candida was found in one patient, and of Enterobacteriaceae in another patient, while small numbers of yeast or Staphylococcus spp. were isolated from the plaque samples of 9 others. 3 out of 4 patients who did not respond to tetracycline treatment had a variety of tetracycline-resistant anaerobic Gram-negative rods present. No correlation was found between increased proportions of tetracycline resistance in the whole bacterial sample and the presence of resistant periodontal pathogens.