Periodontal-disease-associated biofilm: A reservoir for pathogens of medical importance.

The ecological diversity of the periodontal microenvironment may provide suitable conditions for the colonization of species not usually considered members of the oral microbiota. In this investigation, we aimed to determine the prevalence and levels of pathogenic species of medical relevance in the microbiota of individuals with distinct periodontal clinical status. Subgingival biofilm was obtained from patients with periodontal health (H, n = 81), gingivitis (G, n = 55), generalized aggressive (AgP, n = 36) or chronic periodontitis (CP, n = 98), and analyzed for 39 microbial taxa using a checkerboard DNA-DNA hybridization technique. Microbial differences among groups, as well as associations between clinical and microbiological parameters were sought by non-parametric and univariate correlation tests. Neisseria spp., Peptostreptococus anaerobius, Candida albicans, enterobacteria, Pseudomonas aeruginosa, Eubacterium saphenum, Clostridium difficile and Olsenella uli were detected in high mean prevalence and counts in the subgingival microbiota of the study population. Species that were more related to periodontal inflammation and tissue destruction at the patient and site levels included enterobacteria, C. albicans, Neisseria spp., P. aeruginosa, O. uli, Hafnia alvei, Serratia marcescens and Filifactor alocis (p < 0.05). In contrast, Fusobacterium necrophorum, Lactobacillus acidophilus, Staphylococcus aureus and Streptococcus pneumoniae were associated with periodontal health (p < 0.05). Pathogenic species of medical importance may be detected in high prevalence and levels in the periodontal microbiota. Regardless of their role in periodontal health or disease, the periodontal biofilm may be a source for dissemination and development of systemic infections by these pathogenic microorganisms.

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.

Watermelon (Citrullus lanatus) rind flour: Development and characterization of a novel watermelon byproduct.

Watermelon (Citrullus lanatus) is a fruit widely consumed by the Brazilian population; however, its rind is usually discarded, despite its nutritional value. This work aimed to develop a watermelon rind flour (WRF) and a fiber-rich bread. The WRF was submitted to microbiological analysis, proximate composition, antioxidant activity, and a profile of phenolic compounds. Six types of bread were developed: three using WRF (20%, 30%, and 40% of wheat flour replacement) and three control samples (only wheat flour), which were submitted to the same analysis, in addition to colorimetric properties, instrumental texture profile, and sensory evaluation (check-all-that-apply, purchase intent, and acceptance test). A high fiber content was observed in WRF (27.15%). The total concentration of phenolic compounds was 2.38 ± 0.20 mg of gallic acid equivalent per g and benzoic acid was the main compound found (73.50 mg/100 g WRF). The results of the antioxidant capacity through the DPPH method indicated a 30% reduction. The WRF 40% bread had 3.06 g of insoluble fiber, characterizing itself as a source of fiber. In the instrumental texture analysis, it was observed that bread hardness increased significantly as the WRF content increased. The formulation of the bread containing WRF is an alternative for the use of waste, favoring the development of a fiber-source product, with a functional food claim.

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.

Antimicrobial susceptibility and virulence of Enterococcus spp. isolated from periodontitis-associated subgingival biofilm.

BACKGROUND: This study evaluated the prevalence, virulence and antimicrobial susceptibility of enterococci isolated from the subgingival microbiota of patients with different periodontal status. METHODS: Subgingival biofilm was obtained from individuals with periodontal health (PH) (n = 139), gingivitis (n = 103), and periodontitis (n = 305) and cultivated on selective media. Isolated strains were identified by mass spectrometry. Antimicrobial sensitivity was determined by disk diffusion, virulence genes by polymerase chain reaction, and the subgingival microbiota by checkerboard. Differences among groups were assessed by Kruskal-Wallis, Mann-Whitney, and Chi-square tests. RESULTS: Enterococcus spp. were isolated from 7.4% of all samples; 53.7% were Enterococcus faecalis. They were more prevalent in periodontitis (9.8%) and gingivitis (7.8%) than PH (2.2%; P <0.05), but no differences among stages of disease severity were observed. High rates of low susceptibility/resistance (>64%) to at least one antimicrobial were observed. Predominant virulence factors included ace (64.3%), asa (39.3%), and esp (35.7%). Fusobacterium nucleatum was prevalent in the subgingival microbiota of enterococci+ individuals, whereas Dialister pneumosintes was found in low frequency in patients with bopD+ enterococci. Oral streptococci were prevalent (>70%) in patients carrying enterococci susceptible to doxycycline (P <0.05), usually bopD- and esp- (P <0.01). CONCLUSIONS: E. faecalis is increased in periodontitis-associated biofilm. Oral enterococci carry virulence genes and express resistance to some antibiotics commonly used in dentistry, such as ciprofloxacin and erythromycin. Specific subgingival taxa are associated with oral enterococci, suggesting they may interact with species of the dysbiotic periodontitis biofilm, constituting a potential source of factors to tissue destruction, antibiotic resistance dissemination, and poor response to periodontal therapy.

Detection of Helicobacter pylori by polymerase chain reaction in the subgingival biofilm and saliva of non-dyspeptic periodontal patients.

BACKGROUND: Helicobacter pylori has been associated with the development of peptic ulcers and gastric cancer. Although the oral cavity may be a source of transmission, it is unknown whether it acts as a permanent reservoir for this bacterium, particularly in the presence of periodontal disease. The purpose of this study was to evaluate the prevalence of H. pylori by polymerase chain reaction (PCR) in the subgingival biofilm and saliva of subjects with periodontitis. METHODS: Samples were obtained from 56 periodontally healthy subjects and 169 subjects with chronic periodontitis. DNA was extracted from the samples, and the detection of H. pylori was carried out by PCR using the JW22/23 primers. RESULTS: In general, H. pylori was detected in 24% of all samples evaluated. A significantly higher prevalence of H. pylori was observed in subgingival biofilm samples (33.3%) compared to saliva samples (20%) (P <0.05). H. pylori was detected significantly more often in the saliva and subgingival samples from subjects with periodontitis (23.5% and 50%, respectively) compared to samples from periodontally healthy subjects (7.3% and 11.4%, respectively; P <0.05). CONCLUSION: H. pylori was detected frequently in the oral microbiota of subjects with periodontitis, suggesting that periodontal pocketing and inflammation may favor the colonization by this species.

Prevalence of Enterococcus faecalis in subgingival biofilm and saliva of subjects with chronic periodontal infection.

BACKGROUND AND AIMS: Enterococci are increasingly associated with nosocomial and opportunistic infections in humans. The role of the oral cavity as a reservoir for this species is unclear, particularly in the presence of oral infection. This study investigated the prevalence of Enterococcus faecalis in subgingival biofilm and saliva of patients with periodontal disease. METHODS: Samples were obtained from 56 periodontally healthy and 169 chronic periodontitis subjects. DNA was extracted from the samples and detection of E. faecalis was carried out by polymerase chain reaction. RESULTS: In general, E. faecalis was detected in 34.9% of all samples evaluated. No significant difference in the prevalence of this species between subgingival biofilm (34.6%) and saliva (35.1%) samples was observed. E. faecalis was detected significantly more often in saliva and subgingival samples of periodontitis patients (40.5% and 47.8%, respectively) compared to controls (14.6% and 17.1%, respectively; p<0.05). Moreover, significant positive correlations were observed between the presence of E. faecalis and clinical parameters of probing depth, clinical attachment level, bleeding on probing and plaque accumulation (p<0.001). CONCLUSION: The present data showed that E. faecalis is frequently detected in the oral microbiota of periodontitis patients suggesting that periodontal infection may favour the colonization by this species. Close attention should be given to these patients regarding the risk for development of E. faecalis infection in other sites of the body.

Perfil sociodemográfico e situação de saúde dos pacientes expostos ao Césio 137 em Goiânia / Sociodemographic profile and health situation of patients exposed to Cesium 137 in Goiânia

Este trabalho teve por objetivo analisar os aspectos sociodemográficos e da situação de saúde dos indivíduos expostos ao Césio 137, que constam com cadastro ativo no Sistema de Informação ao Radioacidentado (SISRAD) do Centro de Assistência aos Radioacidentados (CARA). Métodos: Trata-se de um estudo descritivo, com análise transversal, realizado no período de outubro de 2019 a fevereiro de 2020 e que contempla dados registrados desde o ano de 1987. O público-alvo da pesquisa consistiu em indivíduos expostos de forma direta e indireta ao Césio 137 na cidade de Goiânia-Goiás, e que se encontram devidamente cadastrados no CARA. Levou-se em consideração aspectos sociodemográficos, a fim de construir o perfil dos pacientes. Realizado esse processo, procedeu-se à classificação e distribuição dos pacientes, através de critérios como a forma de contato e a dose de exposição. O levantamento de dados foi executado pela análise dos prontuários no SISRAD. Resultados e Conclusões: nos grupos mais jovens não foram identificados cânceres precoces ou malformações nos Grupos NGI e NGII, mas existem relatos que conflitam com as declarações de óbitos dos grupos GI e GII, o que trouxe dificuldades para uma análise estratificada e detalhada. As doenças psíquicas, apresentam maior prevalência, seguidas de causas associadas a hipertensão e a diabetes. Verificou-se a importância da análise de saúde dos radioacidentados como ferramenta de planejamento, preparação e avaliação dos grupos de risco

This study aimed to analyze the sociodemographic aspects and the health situation of individuals exposed to Cesium 137, who have an active record in the Information System for Radio Accidents (SISRAD) of the Assistance Center for Radio Accidents (CARA). Methods: This is a descriptive study, with cross-sectional analysis, carried out from October 2019 to February 2020 and which includes data recorded since 1987. The target audience of the research consisted of individuals exposed directly and indirect exposure to Cesium 137 in the city of Goiânia-Goiás, and who are duly registered with the CARA. Sociodemographic aspects were taken into account in order to build the profile of patients. Once this process was completed, the patients were classified and distributed using criteria such as the form of contact and the dose of exposure. Data collection was carried out by analyzing the medical records in SISRAD. Results and Conclusions: In the younger groups, no early cancers or malformations were identified in Groups NGI and NGII, but there are reports that conflict with the death certificates of groups GI and GII, which brought difficulties for a stratified and detailed analysis. Mental illnesses are more prevalent, followed by causes associated with hypertension and diabetes. The importance of analyzing the health of radio-accidents as a tool for planning, preparing and evaluating risk groups was verified

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.

Clinical and microbiological profiles of human immunodeficiency virus (HIV)-seropositive Brazilians undergoing highly active antiretroviral therapy and HIV-seronegative Brazilians with chronic periodontitis.

BACKGROUND: This study compares the periodontal clinical profile and the composition of the subgingival microbiota of human immunodeficiency virus (HIV)-seropositive and HIV-seronegative subjects with chronic periodontitis. METHODS: A total of 172 subjects were distributed into two HIV-seropositive groups (37 chronic periodontitis [H+CP+] and 35 periodontally healthy [H+CP-] individuals) and two HIV-seronegative groups (49 chronic periodontitis [H-CP+] and 51 periodontally healthy [H-CP-] subjects). Subgingival samples were collected from six sites with the deepest probing depth in the periodontitis groups and six random sites in the groups with periodontal health. All HIV-infected patients had undergone highly active antiretroviral therapy (HAART) for at least 2 years. The presence and levels of 33 bacterial species were detected by DNA probes and the checkerboard method. Kruskal-Wallis and Mann-Whitney tests were used to seek for significant differences among and between groups. RESULTS: H-CP+ patients showed significantly more periodontal destruction and inflammation than H+CP+ patients, whereas H+CP- subjects presented a greater percentage of sites with bleeding than H-CP- subjects (P <0.01). Patients who were HIV seronegative showed higher prevalence and levels of most bacterial species than HIV seropositive patients. Periodontal pathogens including Tannerella forsythensis, Porphyromonas gingivalis, Prevotella nigrescens, Eubacterium nodatum, Fusobacterium nucleatum, and Selenomonas noxia were more frequently detected in H-CP+ subjects compared to H+CP+ and controls. In contrast, Enterococcus faecalis and Acinetobacter baumannii were more commonly found in HIV-infected than in non-HIV-infected subjects (P <0.05). CONCLUSION: Putative periodontal pathogens are more prevalent in the subgingival microbiota of HIV-seronegative patients with chronic periodontitis, whereas species not usually associated with periodontitis are detected in higher frequency in HIV-seropositive subjects under HAART.

Effects of non-surgical mechanical therapy on the subgingival microbiota of Brazilians with untreated chronic periodontitis: 9-month results.

BACKGROUND: Mechanical periodontal therapy is the most common treatment of periodontal infections. It is directed primarily towards removing biofilm and calculus from the root surfaces, leading to ecological changes in the subgingival environment. Thus, the purpose of this study was to evaluate the effects of scaling and root planing (SRP) on the subgingival microbiota of Brazilian subjects with untreated chronic periodontitis over a 9-month period. METHODS: Twenty-five untreated chronic periodontitis patients (mean age 43 +/- 5 years; 20% smokers; 45% males) were selected from a Brazilian population. At baseline, probing depth (PD), clinical attachment level (CAL), visible supragingival biofilm (SB), bleeding on probing (BOP), and suppuration (SUP) were measured at six sites/tooth. Subgingival plaque samples were obtained from 10 sites with the deepest PD (> or =5 mm) of each subject and tested for the presence of 25 oral species by DNA probes and the checkerboard technique. Patients received full mouth SRP and oral hygiene instructions. Clinical and microbiological assessments were repeated at 3, 6, and 9 months after therapy. During this period, all patients received maintenance therapy, including supragingival prophylaxis and reinforcement in home care procedures. The clinical and microbiological parameters examined were computed for each subject and at each visit. Differences over time were sought using the Friedman test. RESULTS: Significant reductions in mean CAL and PD (P <0.01), percent of sites with SB (P <0.01), BOP and SUP (P <0.05) were observed during the course of the study. In general, microbial changes were more pronounced for the mean counts than for the frequency of the microorganisms, particularly at 3 months post-therapy. Significant reductions in prevalence and levels were observed for certain periodontal pathogens including P. gingivalis (P <0.05; P <0.01), T. forsythensis (P <0.01), C. rectus (P <0.01), and A. actinomycetemcomitans (P <0.01; P <0.05). Nevertheless, the frequency of A. actinomycetemcomitans increased to baseline values at 9 months after therapy. Treponema ssp. and Prevotella spp. showed a modest decrease in prevalence, whereas marked reductions in their levels were observed. In contrast, the frequency and counts of the suspected pathogens P. micros and F. nucleatum increased after treatment. Species considered beneficial including Actinomyces spp., some oral streptococci, and V. parvula increased in prevalence, although these two last species tended to return to baseline levels at 9 months. CONCLUSION: In Brazilians with untreated chronic periodontitis, SRP led to clinical improvement associated with a decrease of certain periodontal pathogens, and an increase of beneficial species for up to 9 months after therapy.

Endodontic therapy associated with calcium hydroxide as an intracanal dressing: microbiologic evaluation by the checkerboard DNA-DNA hybridization technique.

This study evaluated the predominant microbiota of infected necrotic pulps and the effects of calcium hydroxide therapy on these microorganisms by the checkerboard DNA-DNA hybridization technique. Conventional endodontic therapy associated with calcium hydroxide as intracanal dressing was performed in 12 single-rooted teeth with pulp necrosis and periradicular bone lesion. Samples were collected from the canal at baseline and 14 days after therapy, and the presence of 44 bacterial species was determined by the checkerboard method. Significant differences in the microbiota from baseline to post-therapy were sought by the paired-samples t test. The most prevalent species included F. nucleatum ss. vincentii, C. sputigena, C. ochracea, S. constellatus, V. parvula, P. gingivalis, P. melaninogenica, and S. sanguis. Most of the microorganisms were reduced after treatment, particularly A. gerencseriae, A. israelii, A. naeslundii, C. gingivalis, C. ochracea, P. gingivalis, S. noxia, S. sanguis, and S. oralis (p < 0.05). Conversely, A. actinomycetemcomitans, C. sputigena, and E. corrodens increased in numbers after therapy. These results indicate that conventional endodontic therapy with calcium hydroxide results in the reduction of pathogenic species associated with pulp necrosis. However, its use is limited, because it did not eliminate the whole spectrum of microorganisms.

In vivo evaluation of the effect of essential oil-containing oral strips on salivary bacteria using the checkerboard method.

OBJECTIVE: The aim of this study was to evaluate the antimicrobial effect of essential oil-containing oral strips on different species of the oral microbiota. METHODOLOGY: Saliva samples were collected from 20 subjects with good oral health, diluted and plated onto blood agar medium. The subjects were asked to place the strip (Listerine PocketPaks) on the tongue allowing it to dissolve. After 30 minutes, new saliva samples were collected again and the plates with the samples were incubated under anaerobic conditions at 37 degrees C for seven days. Colony counts (CFU/mL) were determined for each sample. The colonies on the plates were washed with 1 mL of TE buffer, and the bacterial suspensions were processed for the identification of 24 species by DNA probes and the Checkerboard DNA-DNA hybridization method. Differences in total counts, prevalence, and levels of the species evaluated before and after placement of the strips were determined by Wilcoxon sign rank and Chi-square tests. RESULTS: A modest increase in the total bacterial number in saliva from 1.4 x 10(8) to 1.7 x 10(8) bacterial cells was observed 30 minutes after the strip placement, although this change was not significant (p = 0.632). Most of the species reduced in frequency and/or levels, including the pathogens A. actinomycetemcomitans, C. rectus, E. corrodens, Fusobacterium spp., P. intermedia, and S. noxia, as well as the beneficial species A. meyeri, A. georgia, A. gerencseriae, A. odontolyticus, and P. acnes after strip placement. In contrast, A. viscosus, P. melaninogenica, P. gingivalis, P. micros, Streptococcus spp., T. forsythensis, and V. parvula presented an increase in prevalence and/or levels. These changes were not statistically significant after adjusting for multiple comparisons (p > 0.0022). CONCLUSION: The use of the essential oil-containing oral strips resulted in a short-term small increase in the total number of salivary microorganisms. In addition, a not significant decrease of certain periodontopathogens, and an increase in species compatible with oral health were observed.

Long-term evaluation of the antimicrobial susceptibility and microbial profile of subgingival biofilms in individuals with aggressive periodontitis.

This study evaluates the antimicrobial susceptibility and composition of subgingival biofilms in generalized aggressive periodontitis (GAP) patients treated using mechanical/antimicrobial therapies, including chlorhexidine (CHX), amoxicillin (AMX) and metronidazole (MET). GAP patients allocated to the placebo (C, n = 15) or test group (T, n = 16) received full-mouth disinfection with CHX, scaling and root planning, and systemic AMX (500 mg)/MET (250 mg) or placebos. Subgingival plaque samples were obtained at baseline, 3, 6, 9 and 12 months post-therapy from 3-4 periodontal pockets, and the samples were pooled and cultivated under anaerobic conditions. The minimum inhibitory concentrations (MICs) of AMX, MET and CHX were assessed using the microdilution method. Bacterial species present in the cultivated biofilm were identified by checkerboard DNA-DNA hybridization. At baseline, no differences in the MICs between groups were observed for the 3 antimicrobials. In the T group, significant increases in the MICs of CHX (p < 0.05) and AMX (p < 0.01) were detected during the first 3 months; however, the MIC of MET decreased at 12 months (p < 0.05). For several species, the MICs significantly changed over time in both groups, i.e., Streptococci MICs tended to increase, while for several periodontal pathogens, the MICs diminished. A transitory increase in the MIC of the subgingival biofilm to AMX and CHX was observed in GAP patients treated using enhanced mechanical therapy with topical CHX and systemic AMX/MET. Both protocols presented limited effects on the cultivable subgingival microbiota.

Subgingival microbiota of Brazilian subjects with untreated chronic periodontitis.

BACKGROUND: Different periodontopathogenic microbiota have been associated with periodontal diseases in several populations. The present investigation determined the subgingival microbiota of untreated chronic periodontitis Brazilians using the checkerboard DNA-DNA hybridization technique. METHODS: Twenty-five periodontitis patients (mean age, 41 +/- 2; mean probing depth [PD], 3.3 +/- 0.2; mean attachment level [AL], 3.6 +/- 0.2) with no history of previous periodontal therapy and a control group of 14 healthy subjects (mean age, 34 +/- 0.6; mean PD, 1.8 +/- 0.2; mean AL, 1.7 +/- 0.1) were selected. Measurements of PD, AL, bleeding on probing, plaque accumulation, and suppuration were recorded at 6 sites/tooth. Subgingival plaque samples were obtained from 4 sites in each tooth/subject in both groups. The presence and levels of 41 subgingival species were determined in 4,032 plaque samples using whole genomic DNA probes and the checkerboard method. RESULTS: Periodontal pathogens, as well as some unusual species (E. faecalis, E. coli and Bartonella sp.), were detected significantly more often and/or in higher levels in the periodontitis group (P < 0.05). Most species were more frequently detected in interproximal sites. B. forsythus, P. gingivalis, E. nodatum, and F. nucleatum ss vincentii showed a significant positive correlation with mean PD and AL (P < 0.05). CONCLUSIONS: The subgingival microbiota of Brazilians with untreated chronic periodontitis were complex, including high proportions of periodontopathogens commonly found in other populations, as well as some unusual species.

Actinomyces species, streptococci, and Enterococcus faecalis in primary root canal infections.

The purpose of this study was to evaluate the prevalence of Actinomyces species, streptococci, and Enterococcus faecalis in primary root canal infections by using a molecular genetic method. Samples were obtained from 53 infected teeth, of which 27 cases were diagnosed as acute periradicular abscesses. DNA was extracted to evaluate the occurrence of 13 bacterial species by using whole genomic DNA probes and checkerboard DNA-DNA hybridization. Polymerase chain reaction using an ubiquitous bacterial primer was undertaken to check the presence of bacterial DNA in clinical samples. All root canal samples contained bacteria as demonstrated by polymerase chain reaction. The checkerboard DNA-DNA hybridization assay allowed the detection of streptococci in 22.6% of the samples, Actinomyces species in 9.4%, and E. faecalis in 7.5%. The most prevalent species were members of the Streptococcus anginosus group. With regard to the asymptomatic lesions, the most prevalent species were S. intermedius (11.5% of the cases), E. faecalis (11.5%), and S. anginosus (7.7%). S. constellatus was the most prevalent species in pus samples (25.9% of the cases). The other most prevalent species in abscessed teeth were A. gerencseriae (14.8%), S. gordonii (11.1%), S. intermedius (11.1%), A. israelii (7.4%), S. anginosus (7.4%), and S. sanguis (7.4%). S. constellatus was the only species positively associated with acute periradicular abscess (p < 0.01).

Association of T CD4 lymphocyte levels and subgingival microbiota of chronic periodontitis in HIV-infected Brazilians under HAART.

OBJECTIVE: The aim of this study was to determine the subgingival microbiota of HIV-infected patients with chronic periodontitis and different T CD4 lymphocyte levels under HAART. STUDY DESIGN: 64 HIV+ patients (mean age 34.5 +/- 7.3; 75% males) were distributed into Group I: chronic periodontitis (> or = 3 sites with probing pocket depth (PPD) and/or clinical attachment level (CAL) > or = 5 mm); and Group II: periodontal health (no sites with PPD > 3 mm and/or CAL > 4 mm). All subjects received conventional periodontal therapy. Periodontal clinical parameters were evaluated at 6 sites/tooth in all teeth at baseline and 4 months after therapy. The levels of T CD4 were obtained from the patient's medical record. Subgingival plaque samples were taken from the 6 sites with the largest pocket depth in each subject of Group I, and 6 randomly selected sites in subjects of Group II. The presence of 22 subgingival species was determined using the checkerboard DNA-DNA hybridization method. Significant microbiological differences within and among groups were sought using Wilcoxon signed-rank and Mann-Whitney tests, respectively. Relationships between T CD4 levels and microbiological parameters were determined using Kruskal-Wallis test. RESULTS: Sixty-one percent of the HIV-infected patients represented AIDS cases, although 69% of them were periodontally healthy. The T CD4 lymphocyte mean level was 333 cells/mm3 and viral load was 12,815 +/- 24,607 copies/mm3. Yet, the prevalence of chronic periodontitis was relatively low (36%). Several periodontal pathogens, in particular T. forsythensis (P < .05), were more prevalent in HIV-positive patients with periodontitis than in HIV-positive subjects with periodontal health. Most of the species decreased in frequency after therapy, particularly P. gingivalis (P < .05). E. faecalis and F. nucleatum were significantly more prevalent in the subgingival microbiota of patients with chronic periodontitis and lower levels of T CD4 (P < .05), while beneficial species tended to be more frequently detected in individuals with T CD4 counts over 500 cells/mm3. CONCLUSION: The subgingival microbiota of HIV-infected patients with chronic periodontitis include a high prevalence of classical periodontal pathogens observed in non-infected individuals. Furthermore, the severe immunosuppression seems to favor the colonization by these species, as well as by species not commonly found in the subgingival microbiota.

Prevalence of Pseudomonas aeruginosa and Acinetobacter spp. in subgingival biofilm and saliva of subjects with chronic periodontal infection.

P. aeruginosa and Acinetobacter spp. are important pathogens associated with late nosocomial pneumonia in hospitalized and institutionalized individuals. The oral cavity may be a major source of these respiratory pathogens, particularly in the presence of poor oral hygiene and periodontal infection. This study investigated the prevalence of P. aeruginosa and Acinetobacter spp. in subgingival biofilm and saliva of subjects with periodontal disease or health. Samples were obtained from 55 periodontally healthy (PH) and 169 chronic periodontitis (CP) patients. DNA was obtained from the samples and detection of P. aeruginosa and Acinetobacter spp. was carried out by multiplex and nested PCR. P. aeruginosa and Acinetobacter spp. were detected in 40% and 45% of all samples, respectively. No significant differences in the distribution of these microorganisms between men and women, subgingival biofilm and saliva samples, patients ≤ 35 and > 35 years of age, and smokers and non-smokers were observed regardless periodontal status (p > 0.05). In contrast, the frequencies of P. aeruginosa and Acinetobacter spp. in saliva and biofilm samples were significantly greater in CP than PH patients (p < 0.01). Smokers presenting P. aeruginosa and high frequencies of supragingival plaque were more likely to present CP than PH. P. aeruginosa and Acinetobacter spp. are frequently detected in the oral microbiota of CP. Poor oral hygiene, smoking and the presence of P. aeruginosa are strongly associated with periodontitis.

Prevalência de bactérias Gram-positivas em infecção do trato urinário / Prevalence of Gram-positive bacterias in urinary tract infection

Objetivo: Identificar a prevalência de bactérias patogênicas Gram-positivas causadoras de ITU, analisando a faixa etária e o gênero mais acometido. Métodos: Este estudo analisou 24.529 prontuários de pacientes ambulatoriais, realizado no Laboratório Clínico da Pontifícia Universidade Católica do Goiás (LC/PUC Goiás), no período de janeiro de 2011 a dezembro de 2017. Foram incluídos neste estudo pacientes de ambos os gêneros e de todas as idades. Os dados foram coletados a partir do livro de registros de amostras específicas para a seção de microbiologia e selecionados os resultados de cultura positivos para bactérias Gram-positivas. Resultados: Das 24.529 uroculturas analisadas, 21,2% (804) apresentaram diagnóstico positivo para ITU por bactérias Gram-positivas. Dentre estas, 85,0% eram do gênero feminino, sendo que os estafilococos coagulase negativa foram os agentes patogênicos de maior prevalência (57,8%). Conclusão: A infecção do trato urinário foi mais frequente em mulheres na faixa etária de 19 a 40 anos, apresentando os estafilococos coagulase negativa os principais uropatógenos associados à infecção, seguidos do Staphylococcus aureus. A identificação de bactérias Gram-positivas tem se mostrado de extrema importância, principalmente para o tratamento correto dos pacientes.

Objective: To identify the prevalence of Gram-positive pathogenic bacteria that cause UTI, analyzing the age group and the most affected genus. Methods: A retrospective study was carried out, in which 24,529 records of outpatients were analyzed, performed at the Clinical Laboratory of the Pontifical Catholic University of Goiás (LC/PUC Goiás) from January 2011 to December 2017. Both genders and of all ages. Data were collected from the log book of samples specific to the microbiology section and selected the culture results positive for Gram positive bacteria. Results: Of the 24,529 urocultures analyzed, 21,2%(804) presented a positive diagnosis for UTI for Gram positive bacteria. Of these, 85.0% were female, and coagulase negative staphylococci were the most prevalent pathogens (57.8%). Conclusion: Urinary tract infection was more frequent in women aged of 19-40 years, with coagulase-negative Staphylococcus being the main uropathogens associated with infection, followed by Staphylococcus aureus. The identification of Gram-positive bacteria has been show to be extremely important, especially for the correct treatment of patients.