This study assessed the antimicrobial activity of a new bioactive glass-ceramic (Biosilicate®) against anaerobic, microaerophilic, and facultative anaerobic microorganisms. Evaluation of the antimicrobial activity was carried out by three methods, namely agar diffusion, direct contact, and minimal inhibitory concentration (MIC). For the agar diffusion technique, bio glass-ceramic activity was observed against various microorganisms, with inhibition haloes ranging from 9.0 ± 1.0 to 22.3 ± 2.1 mm. For the direct contact technique, Biosilicate® displayed activity against all the microorganisms, except for S. aureus. In the first 10 min of contact between the microorganisms and Biosilicate®, there was a drastic reduction in the number of viable cells. Confirming the latter results, MIC showed that the Biosilicate® inhibited the growth of microorganisms, with variations between ≤ 2.5 and 20 mg/ml. The lowest MIC values (7.5 to ≤ 2.5 mg/ml) were obtained for oral microorganisms. In conclusion, Biosilicate® exhibits a wide spectrum of antimicrobial properties, including anaerobic bacteria.