Although specific subgingival microflora are being associated with active tissue destruction in periodontitis, little information exists on the relationship between subgingival flora and histological evidence of tissue destruction. The present study was undertaken to explore these relationships during experimental marginal periodontitis in squirrel monkeys. In 4 squirrel monkeys, experimental periodontitis was induced around bicuspids and molars using silk ligatures. The buccal part of the subgingival ligature was removed for bacterial sampling 3, 7, and 14 days after ligature placement. Dark-field microscopy was used to quantify motile forms, spirochetes, straight and curved rods, filaments, cocci and fusiforms. Subgingival plaque prior to periodontitis induction was dominated by cocci, but fusiforms and straight rods were also present. Spirochetes, filaments, curved rods and motile forms were absent. 3 days after induction of periodontitis, there were marked increases in spirochetes, motile forms, filaments, curved and straight rods. The proportions of coccoids and fusiforms decreased. The bacterial proportions stayed at approximately the same levels through 7 and 14 days. Histometric quantification of periodontal destruction showed loss of connective tissue attachment and crestal alveolar bone. A positive correlation was found between periodontal tissue destruction and both total number of subgingival bacteria and spirochetes.