Prevalence of Enterococcus species in adults with periodontal health or with periodontitis: a systematic review.

The aim of this study was to evaluate the prevalence of Enterococcus species in the mouth of adults with periodontal health and periodontitis. A systematic search was made in databases in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. The search for articles was conducted in Medline/PubMed, Latin American and Caribbean Health Sciences Literature Database (LILACS), Cochrane Library, Scopus, Embase, Web of Science databases and in the System of Information on Grey Literature in Europe (SINGLE) and included articles published in English up to April 25th, 2021. Observational studies in humans with and without periodontitis were evaluated to identify the prevalence of Enterococcus species. Articles that met the inclusion criteria were analyzed and classified to determine the quality rating in good, fair, and poor. A new detailed checklist for quality assessment was developed based on the information required for applicable data extraction in reviews. The study design, sample size, demographic data, periodontal clinical parameters, microbial analysis method, biological sample, prevalence of Enterococcus spp., and correlations with periodontal clinical parameters were assessed. After screening and full-text reading, 8 articles met the inclusion criteria. All selected studies showed a significantly higher prevalence of Enterococcus spp. in patients with periodontitis compared with periodontally healthy patients. Thus, the present systematic review suggests that the prevalence of Enterococcus faecalis in the mouth of periodontitis individuals is higher than that of periodontally healthy individuals.

Antimicrobial resistance and virulence of subgingival staphylococci isolated from periodontal health and diseases.

The dysbiotic biofilm of periodontitis may function as a reservoir for opportunistic human pathogens of clinical relevance. This study explored the virulence and antimicrobial susceptibility of staphylococci isolated from the subgingival biofilm of individuals with different periodontal conditions. Subgingival biofilm was obtained from 142 individuals with periodontal health, 101 with gingivitis and 302 with periodontitis, and cultivated on selective media. Isolated strains were identified by mass spectrometry. Antimicrobial susceptibility was determined by disk diffusion. The mecA and virulence genes were surveyed by PCR. Differences among groups regarding species, virulence and antimicrobial resistance were examined by Chi-square, Kruskal-Wallis or Mann-Whitney tests. The overall prevalence of subgingival staphylococci was 46%, especially in severe periodontitis (> 60%; p < 0.01). S. epidermidis (59%) and S. aureus (22%) were the predominant species across groups. S. condimenti, S. hominis, S. simulans and S. xylosus were identified only in periodontitis. High rates of resistance/reduced sensitivity were found for penicillin (60%), amoxicillin (55%) and azithromycin (37%), but multidrug resistance was observed in 12% of the isolates. Over 70% of the mecA + strains in periodontitis were isolated from severe disease. Higher detection rates of fnB + isolates were observed in periodontitis compared to health and gingivitis, whereas luxF/luxS-pvl + strains were associated with sites with deep pockets and attachment loss (p < 0.05). Penicillin-resistant staphylococci is highly prevalent in the subgingival biofilm regardless of the periodontal status. Strains carrying virulence genes related to tissue adhesion/invasion, inflammation and cytotoxicity support the pathogenic potential of these opportunists in the periodontal microenvironment.

Prevalence of Enterococcus species in adults with periodontal health or with periodontitis: a systematic review

Abstract The aim of this study was to evaluate the prevalence of Enterococcus species in the mouth of adults with periodontal health and periodontitis. A systematic search was made in databases in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. The search for articles was conducted in Medline/PubMed, Latin American and Caribbean Health Sciences Literature Database (LILACS), Cochrane Library, Scopus, Embase, Web of Science databases and in the System of Information on Grey Literature in Europe (SINGLE) and included articles published in English up to April 25th, 2021. Observational studies in humans with and without periodontitis were evaluated to identify the prevalence of Enterococcus species. Articles that met the inclusion criteria were analyzed and classified to determine the quality rating in good, fair, and poor. A new detailed checklist for quality assessment was developed based on the information required for applicable data extraction in reviews. The study design, sample size, demographic data, periodontal clinical parameters, microbial analysis method, biological sample, prevalence of Enterococcus spp., and correlations with periodontal clinical parameters were assessed. After screening and full-text reading, 8 articles met the inclusion criteria. All selected studies showed a significantly higher prevalence of Enterococcus spp. in patients with periodontitis compared with periodontally healthy patients. Thus, the present systematic review suggests that the prevalence of Enterococcus faecalis in the mouth of periodontitis individuals is higher than that of periodontally healthy individuals.

Prevalence and antimicrobial susceptibility of Gram-negative bacilli in subgingival biofilm associated with periodontal diseases.

BACKGROUND: This cross-sectional study aimed to determine the prevalence and antimicrobial susceptibility of Gram-negative bacilli (GNB) isolated from subgingival biofilm of individuals with different periodontal conditions. METHODS: Subgingival biofilm was obtained from 362 individuals with periodontal health (PH) (n = 83), gingivitis (n = 74), and periodontitis (n = 205), cultivated in broth and selective media. Isolated strains were identified by mass spectrometry. Antimicrobial susceptibility was determined by the Clinical and Laboratory Standards Institute disk diffusion guidelines. Production of extended-spectrum beta-lactamase (ESBL) and carbapenemases were evaluated by double disk synergy test and spectrophotometric detection of imipenem hydrolysis, respectively. ESBL and carbapenemase encoding genes were surveyed by Polymerase chain reaction (PCR). Differences among groups were examined by Chi-square, Kruskal-Wallis or Mann-Whitney tests. RESULTS: GNB were isolated from 36.2% of all subgingival biofilm samples, with a significantly greater prevalence and species diversity (P < 0.001) in patients with periodontitis (45.9%) compared with individuals with PH (24.1%) and gingivitis (22.9%). Pseudomonas aeruginosa (27.5%), Enterobacter cloacae (16.8%), and Enterobacter asburiae (10.7%) were the most predominant species. Resistance/reduced sensitivity to at least 1 antimicrobial was detected in 60% of the strains, but only 4.6% were multidrug resistant. Serratia marcescens, E. cloacae, and Enterobacter kobei presented high rates of intrinsic resistance (>40%) to amoxicillin-clavulanate and first/second-generations of cephalosporins. One strain of Klebsiella pneumoniae isolated from periodontitis was resistant to imipenem, but no ESBL encoding genes or ESBL phenotype was detected. CONCLUSION: High prevalence and diversity of GNB, with low susceptibility to ß-lactams are observed in the subgingival microbiota associated with periodontitis.

Prevalência, susceptibilidade antimicrobiana e virulência de bacilos Gram-negativos e enterococos isolados do biofilme subgengival associado à doenca periodontal' / Prevalence, antimicrobial susceptibility and virulence of Gram-negative bacilli and enterococci isolated from subgingival biofilm associated with periodontal disease

Objetivos: A cavidade oral pode atuar como reservatório de vários patógenos de importância clínica, incluindo bacilos Gram-negativos (BGN) e Enterococcus spp. Essas espécies podem ainda aumentar em uma condição disbiótica, como ocorre nas doenças periodontais. Assim, este estudo teve como objetivo determinar a prevalência, susceptibilidade antimicrobiana e presença de fatores de virulência de BGN e enterococos isolados do biofilme subgengival de indivíduos com diferentes condições periodontais, correlacionando esses achados com parâmetros clínicos e composição da microbiota subgengival. Métodos: Na análise dos BGN, amostras de biofilme subgengival foram obtidas de indivíduos com Saúde Periodontal (SP, n=81), Gengivite (G, n=74) e Periodontite (P, n=207); para Enterococcus spp., amostras foram coletadas de 139 indivíduos com SP, 103 com G e 305 com P. As amostras foram cultivadas em meio seletivo e as colônias isoladas e identificadas por MALDI-ToF. A susceptibilidade antimicrobiana foi determinada por disco-difusão (CLSI); já os genes de virulência de enterococos e genes codificadores de ESBL e carbapenemases em BGN foram pesquisados por PCR. A produção de ESBL e carbapenemases por BGN foi avaliada pelo teste de sinergia de disco duplo e hidrólise de imipenem por espectrofotometria, respectivamente. A microbiota subgengival desses indivíduos foi determinada pelo Checkerboard. Diferenças entre os grupos foram avaliadas pelos testes de KruskalWallis, Mann-Whitney e Qui-quadrado. Resultados: BGN foram isolados em 36.2% das amostras, com maior prevalência (p<0,001) em pacientes com P (46.4%) em comparação com SP (22.2%) e G (22.9%). Pseudomonas aeruginosa (27.5%), Enterobacter cloacae (16.8%) e Enterobacter asburiae (10.7%) foram as espécies mais predominantes. Resistência/sensibilidade reduzida a ≥ 1 antimicrobiano foi encontrada em 60% dos BGN, mas apenas 4.6% eram multirresistentes. Altas taxas de resistência (>40%) foram observadas na família Enterobacteriacea para cefoxitina, cefalotina, amoxicilina- clavulanato e cefazolina. Uma única cepa de K. pneumoniae apresentou resistência/sensibilidade reduzida ao imipenem, embora o fenótipo ESBL e a detecção dos genes codificadores de beta-lactamases foram negativos. Enterococcus spp. foram isolados em 7.4% de todas as amostras, 53.7% eram E. faecalis. Essas espécies foram mais predominantes na P (9.8%) e G (7.8%) do que na SP (2.2%, p< 0,05); entretanto não houve correlação com os níveis de gravidade da P. Altas taxas de resistência/susceptibilidade reduzida foram observadas para ciprofloxacina, eritromicina e rifampicina. Os fatores de virulência mais predominantes incluíram ace, asa e esp, todos relacionados à formação de biofilme e colonização. F. nucleatum foi mais prevalente na microbiota de indivíduos enterococos +. Por outro lado, Dialister pneumosintes foi pouco detectado em indivíduos portadores de enterococos bopD+. Estreptococos orais foram prevalentes (>70%) na microbiota de pacientes que apresentavam enterococos suceptíveis à doxiciclina (p<0,05), frequentemente bopD- e esp- (p<0,01). Conclusão: Uma prevalência elevada de BGN da família Enterobacteriacea com resistência a cefalosporinas e penicilina é observada na microbiota subgengival de indivíduos com P. Enterococcus spp., principalmente E. faecalis são pouco frequentes na microbiota subgengival associada à SP, porém aumentam significativamente nas doenças periodontais. Os mesmos apresentam diversos genes de virulência compatíveis com destruição tecidual, bem como resistência a antimicrobianos de uso na clínica periodontal, o que pode limitar uma resposta terapêutica favorável. (AU)

Background/Aim: The oral cavity can act as a reservoir for several pathogens of clinic importance, including Gram-negative bacilli (GNB) and Enterococcus spp. These species may increase even more in a dysbiotic condition as seen in periodontal diseases. Thus, this study aimed to determine the prevalence, antimicrobial susceptibility and virulence factors of GNB and enterococci isolated from subgingival biofilm of individuals with different periodontal conditions, correlating these findings with clinical parameters and the composition of the subgingival microbiot. Methods: For GNB analysis, subgingival biofilm was obtained from individuals with periodontal health (PH, n=81), gingivitis (G, n=74) and periodontitis (P, n=207), whereas for enterococci isolation samples were taken from 139 patients with PH, 103 with G, and 305 with P. Samples were cultivated in selective media and isolated colonies were identified by MALDI-ToF. Antimicrobial susceptibility was determined by CLSI disk diffusion, whereas virulence genes by PCR. Production of ESBL and carbapenemases were evaluated by double disk synergy test and spectrophotometric detection of imipenem hydrolysis, respectively, and ESBL and carbapenemase encoding genes were surveyed by PCR. The subgingival microbiota was determined by checkerboard. Differences among groups were examined by Chi-square, Kruskal-Wallis or Mann-Whitney tests. Results: GNB were isolated from 36.2% of all samples, with a significantly greater prevalence (p<0.001) in P patients (46.4%) compared to PH (22.2%) and G (22.9%). Pseudomonas aeruginosa (27.5%), Enterobacter cloacae (16.8%) and Enterobacter asburiae (10.7%) were the most predominant species. Resistance/reduced sensitivity to ≥ 1 antimicrobial was found in 60% of GNB, but only 4.6% were multidrug resistant. High resistance rates (>40%) were seen in the Enterobacteriaceae family for cefoxitin, cephalotin, amoxicillin-clavulanate, and cefazolin. One strain of K. pneumoniae showed resistance/reduced sensitivity to imipenem, although the ESBL-phenotype and PCR targeting beta-lactamase encoding genes were negative. Enterococcus spp. were isolated from 7.4% of all samples; 53.7% were E. faecalis. Enterococci were more predominant in P (9.8%) and G (7.8%) samples than PH (2.2%; p<0.05), however there were no associations with distinct levels of disease severity. High rates of low susceptibility/resistance were seen for ciprofloxacin, erythromycin and rifampicin. Predominant virulence factors included ace, asa and esp, all related to colonization and biofilm formation. F. nucleatum was prevalent in the microbiota of enterococci+ individuals. In contrast, lower frequency of Dialister pneumosintes was found in patients carrying bopD+ enterococci. Oral streptococci were prevalent (>70%) in the microbiota of patients carrying enterococci susceptible to doxycycline (p<0.05), which were also frequently bopD- and esp- (p<0.01). Conclusion: A high prevalence of GNB of the Enterobacteriacea family, resistant to cephalosporins and penicillins is observed in the subgingival microbiota of patients with P, Enterococcus spp., mainly E.faecalis are not commonly detected in the healthy-related subgingival microbiota, however their frequency increases significantly in patients with periodontal diseases. These species carry several genes related to tissue destruction, as well as resistance to antimicrobials routinely used in the periodontal clinic, which may hinder a successful therapeutic response. (AU)

Lack of adjunctive effect of 0.1% sodium hypochlorite mouthwash combined to full-mouth ultrasonic debridement on supragingival plaque, gingival inflammation, and subgingival microbiota: A randomized placebo-controlled 6-month trial.

To test the adjunctive effect of 0.1% sodium hypochlorite (NaOCl) mouthwash combined to full-mouth ultrasonic debridement (FMUD) on reducing supragingival plaque, gingival inflammation, and microbial pathogens. In this 6-month double-blinded randomized clinical trial, individuals with gingivitis were assigned to test (n = 16) or placebo group (n = 16) and received FMUD followed by rinsing with 0.1% NaOCl (test) or distilled water (placebo), respectively, twice a day for 1 month. Full-mouth periodontal examination was performed at baseline, 1, 3, and 6 months posttherapy, and subgingival plaque samples were obtained at the same time points and analysed for their composition by checkerboard. Differences between groups over time were examined by Student t test, Mann-Whitney, generalized linear model, and Friedman and chi-square tests. Both therapeutic protocols resulted in significant clinical improvement in periodontal parameters over time, except for probing depth and attachment level, which had a slight mean increase of 0.2 mm (p < .01). No significant differences between groups were observed for any clinical parameter (p > .05). Most species (>65%) decreased similarly in levels in both groups over time. Significant reductions in the microbial complexes were seen mainly at 1 and 3 months, but they returned to baseline levels in both groups, except for the red and yellow complexes, and other oral species, which were kept in low levels at 6 months (p < .05). A 0.1% NaOCl mouthwash did not provide additional benefits to FMUD in reducing supragingival plaque, gingivitis, and/or microbial pathogens.

Efeito adjunto da solução de hipoclorito de sódio à 0,1% no controle do biofilme dental, gengivite e microbiota oral: estudo clínico randomizado, duplo-cego e controlado / Adjunct effect of 0. 1% sodium hypochlorite solution in the control of dental biofilm, gingivitis and oral microbiota: a randomized, double-blind, controlled clinical trial

Pouca informação existe sobre o uso de enxaguatório bucal contendo Hipoclorito de sódio (NaOCl) como uma terapia alternativa adjuvante para o controle químico do biofilme dental. O presente estudo clínico randomizado, duplo cego e controlado com placebo teve por objetivo avaliar, por um período de 6 meses, a eficácia da solução de NaOCl à 0,1%, utilizada como antisséptico bucal coadjuvante à profilaxia periodontal mecânica, na redução do biofilme supragengival, inflamação gengival, e dos níveis de patógenos periodontais e oportunistas.32 indivíduos diagnosticados com gengivite foram recrutados da Clínica de Periodontia da Universidade Federal do Rio de Janeiro, sendo os mesmos randomizados e alocados em dois grupos terapêuticos. O Grupo placebo (C, n=16) foi submetido à profilaxia periodontal ultra-sônica seguida do uso de água destilada como enxaguante bucal por 1 mês. O Grupo teste (T, n=16) foi submetido à profilaxia seguida do uso do NaOCl à 0,1% como enxaguante. O exame clínico periodontal, incluindo medidas clínicas de profundidade de sondagem (PS), nível clínico de inserção (NCI), sangramento à sondagem (SAS), sangramento gengival (IG), placa supragengival (IP) e cálculo (CA), foi realizado por um examinador calibrado no início do estudo (pré-terapia), 1, 3 e 6 meses pós-terapia. Amostras de saliva e de biofilme subgengival foram obtidas de cada paciente nesses diferentes tempos de avaliação. A composição da microbiota salivar e periodontal foi determinada pelo método do checkerboard. Diferenças entre os grupos em relação aos parâmetros clínicos e à microbiota ao longo do tempo foram avaliadas pelos testes do Qui-quadrado, Student T, GLM, Friedman e Mann-Whitney. Ambos os protocolos terapêuticos resultaram em melhora clínica significativa nos parâmetros periodontais após tratamento, com exceção do aumento clinicamente irrelevante na PS e NCI (teste de Friedman, p<0,01).No entanto, não houve diferença significativa entre os grupos (teste GLM, p>0,05). A maioria da espécies avaliadas na saliva apresentou um aumento em níveis, porém >67% das espécies no biofilme subgengival reduziram em ambos os grupos terapêuticos ao longo dos 6 meses pós-terapia. Reduções significativas nos níveis dos complexos microbianos foram observadas particularmente 1 e 3 meses após tratamento. Entretanto, aos 6 meses esses níveis retornaram a valores da pré-terapia, exceto para os complexos vermelho e amarelo, e outras espécies orais que se mantiverem em níveis baixos pós-terapia em ambos os grupos (p<0.05, GLM). O uso do NaOCl a 0.1% como enxaguante bucal não apresentou um efeito benéfico adicional à profilaxia periodontal ultra-sônica na redução do biofilme supragengival, gengivite e níveis de patógenos orais e oportunistas (AU)

Limited data are available regarding the use of a Sodium Hypochlorite (NaOCl) mouthwash as an alternative adjuvant therapy for dental plaque chemical control. This randomized, double-blinded, placebo-controlled study aimed to evaluate for 6 months the effectiveness of a 0.1% NaOCl solution used as an antiseptic oral rinse adjunct to mechanical periodontal prophylaxis on reducing supragingival biofilm, gingival inflammation, as well as the levels of oral pathogens and opportunists. 32 individuals diagnosed with gingivitis were recruited from the Periodontics Clinic at the Federal University of Rio de Janeiro, randomized and allocated into 2 therapeutics groups. The Placebo group (C, n=16) received ultrassonic periodontal prophylaxis followed by the use of sterile distilled water as mouthwash for 1 month, whereas the Test group (T, n=16) had prophylaxis followed by the use of 0.1% NaOCl as mouthwash. Full-mouth periodontal examination including measurements of probing depth (PD), clinical attachment level (CAL), bleeding on probing (BOP), gingival bleeding (GI), supragingival plaque (PI) and calculus (CA) was carried out by one trained examiner at baseline (pre-therapy), 1, 3 and 6 months pos-therapy. Saliva and subgingival biofilm samples were obtained from each patient at the same follow up time points. The composition of the salivary and periodontal microbiota was determined by the checkerboard method. Significance of differences between groups for clinical and microbiological parameters over time were sought by Chi-square, Student T, GLM, Friedman and Mann-Whitney tests. Both therapeutic protocols resulted in significant clinical improvement in periodontal parameters over time, except for PD and CAL which presented a slight increase (Friedman test, p<0.01).However, no significant differences between groups were observed for these parameters (GLM, p>0.05).Most species in saliva presented an increase in mean counts, whereas >67% of the species in the subgingival biofilm decreased in both therapeutic groups over time. Significant reductions on the microbial complexes levels were observed mainly at 1 and 3 months pos-therapy. However, at 6 months, these complexes rebounded to pre-therapy counts, except for the red and yellow complexes, and other oral species which were maintained at low levels after treatment in both groups (p<0.05, GLM). The use of 0.1% NaOCl as an oral rinse adjunctive to mechanical periodontal prophylaxis did not provide additional benefits on reducing the supragingival biofilm, gingivitis and levels of microbial pathogens (AU)