Recently, dental implants have experienced increasing demand as one of the most effective, permanent and stable ways for replacing missing teeth. However, peri-implant diseases that are multispecies plaque-based infections may ultimately lead to implant failure (i.e., late peri-implantitis). Therefore, the present study aims to detect the microbial diversity of subgingival plaque in peri-implantitis cases (N = 30) by comparing with healthy implants (N = 34) using culture-based identification methods, including VITEK 2 system. An increase in microbial diversity (29 species along with 1 and 7 isolates, which were classified as a genus and unidentified species, respectively) were observed in subgingival sites of diseased implants dominated by Gram negative enteric bacilli compared with healthy implants (21 species with 2 at genus level) with the majority of Grampositive lactic acids species. Our results showed significant differences in the mean age between healthy (53.14±11.34) and diseased implants (61.9±9.71).