| Summary: | Periodontal disease is a relevant oral disease in dogs and nisin–biogel has been previously proposed to be used in its control. <i>Enterococci</i>, as inhabitants of the oral cavity with a high genetic versatility, are a reliable bacterial model for antimicrobial studies. Our goal was to evaluate the in vivo influence of the long-term dental application of the nisin–biogel on the virulence and antimicrobial signatures of canine oral <i>enterococci</i>. Twenty dogs were randomly allocated to one of two groups (treatment group—TG with nisin–biogel dental application, or control group—CG without treatment) and submitted to dental plaque sampling at day 0 and after 90 days (T90). Samples were processed for <i>Enterococcus</i> spp. isolation, quantification, identification, molecular typing and antimicrobial and virulence characterization. From a total of 140 <i>enterococci</i>, molecular typing allowed us to obtain 70 representative isolates, mostly identified as <i>E. faecalis</i> and <i>E. faecium</i>. No significant differences (<i>p</i> > 0.05) were observed in the virulence index of the isolates obtained from samples collected from the TG and CG at T90. At T90, a statistically significant difference (<i>p</i> = 0.0008) was observed in the antimicrobial resistance index between the isolates from the TC and CG. Oral <i>enterococci</i> were revealed to be reservoirs of high resistant and virulent phenotypes.
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