β-lactamase production and antimicrobial susceptibility of subgingival bacteria from refractory periodontitis

This study assessed the extent of beta-lactamase-producing bacteria in subgingival plaque samples obtained from 25 patients with refractory marginal periodontitis in the USA. beta-Lactamase-positive isolates were characterized using commercial diagnostic kits and partial sequencing of the 16S rRNA gene. The susceptibilities to different antimicrobial agents were tested and, in addition, the isolates were screened for the presence of extended spectrum beta-lactamases (ESBLs). beta-lactamase-producing bacteria were detected in 18 (72%) patients. The most prominent beta-lactamase-producing organisms belonged to the anaerobic genus Prevotella. Other enzyme-producing anaerobic strains were Fusobacterium nucleatum, Propionibacterium acnes and Peptostreptococcus sp. Facultative bacteria, such as Burkholderia spp., Ralstonia pickettii, Capnocytophaga spp., Bacillus spp., Staphylococcus spp. and Neisseria sp., were also detected among the enzyme-producers. Minimum inhibitory concentrations (MICs) of ampicillin and amoxicillin were in the range 1.5-256 mug/ml and 4-256 mug/ml, respectively, for the isolates of the Prevotella species. All Prevotella isolates were susceptible to amoxicillin/clavulanate and metronidazole, but they showed variable resistance to tetracyclines. Two of the Prevotella isolates had high MICs of cefotaxime and ceftazidime. ESBL activity was not detected in any of the beta-lactamase-producing isolates by the Etest method. Thus, our study demonstrated a wide variety of beta-lactamase-producing bacteria that may play a role in refractory periodontal disease.