Several studies have tried to associate the presence of different pathogens with the onset and progression of periodontitis, reporting a wide variety of results from different populations and environments. The aim of this study was to determine the main periodontal pathogens present in the subgingival biofilm of Dominican patients with periodontitis, by using specific microbiological culturing techniques. Periodontitis patients were selected after a full­mouth periodontal evaluation, and assigned to different periodontitis groups based on percentage of affected locations. Subgingival samples were collected and analyzed by means of specific culture techniques. Anaerobic counts, frequency of detection and proportions of target pathogens were calculated. Variables were analyzed by means of Student’s T­test or chi­square test. Twenty­nine subjects were recruited, of whom 17 were diagnosed with generalized periodontitis (GenP) and 12 with localized periodontitis (LocP). The most prevalent bacterial species in both groups was Prevotella intermedia (94.1% in GenP and 91.7% in LocP), followed by Porphyromonas gingivalis (88.2% in GenP and 83.3% in LocP). Total microbiota in subgingival samples was 1.3 x107 colony­forming units (CFU)/mL (standard deviation, SD=1.5 x107) and 9.6x106 CFU/mL (SD=1.1 x107) in GenP and LocP subjects, respectively, though differences were not statistically significant (p=0.222). The highest counts were observed for P. gingivalis in both groups, with mean concentration 2.5x106 CFU/mL (6.1x106) in GenP and 2.9x106 CFU/mL (5x106) in LocP, with no statistically significant difference (p=0.879). These results suggest that relevant periodontal pathogens are found with diversity and abundance in the subgingival microbiota of adult Dominican patients with periodontitis.