It has been reported in the literature that certain species of bacteria (periodontopathogens) present in the subgingival plaque are associated with destructive periodontal disease. The purpose of this study was to investigate and compare the presence and proportional distribution of periodontopathogens in the subgingival plaque of adult crowded versus non-crowded dental regions. Thirty adult patients with anterior dental crowding were selected from the Orthodontic Clinic of the University of Pennsylvania. After orthodontic records were taken and the periodontal examination was performed, subgingival plaque samples were collected from crowded (experimental) and contralateral non-crowded regions (control) of each patient. The presence of 9 periodontopathic species, Actinobacillus actinomycetemcomitans, Prevotella intermedia, Eikenella corrodens, Campylobacter rectus, Capnocytophaga species, Fusobacterium species, Peptostreptococcus micros, Porphyromonas gingivalis, and Bacteroides forsythus, was determined using culture and immunofluorescence techniques. The bacterial morphotype was also determined by the use of dark-field microscopy. It was found that supragingival plaque accumulation in crowded regions was significantly greater than in non-crowded regions. Analysis of the bacteria showed that the samples from crowded regions consistently contained more species of periodontopathogens than the samples from non-crowded regions. Morphologically, more spirochetes and motile rods were present in the crowded-region samples. In terms of the presence of individual periodontopathogens, Fusobacterium species, Capnocytophaga species, C rectus, and P micros were significantly more common in the crowded samples than in the non-crowded samples. It was concluded that: (1) more plaque accumulated in crowded areas; (2) more species of periodontopathogens were present in the subgingival plaque of crowded regions; (3) morphologically, more spirochetes and motile rods were present in crowded areas; and (4) Fusobacterium species, Capnocytophaga species, C rectus, and P micros were present more often in crowded areas than in non-crowded areas (P < 0.05)