Real-Time PCR Method as Diagnostic Tool for Detection of Periodontal Pathogens in Patients with Periodontitis.

The most common type of periodontal disease is chronic periodontitis, an inflammatory condition caused by pathogenic bacteria in subgingival plaque. The aim of our study was the development of a real-time PCR test as a diagnostic tool for the detection and differentiation of five periodontopathogenic bacteria, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Prevotella intermedia, and Treponema denticola, in patients with periodontitis. We compared the results of our in-house method with the micro-IDent® semiquantitative commercially available test based on the PCR hybridization method. DNA was isolated from subgingival plaque samples taken from 50 patients and then analyzed by both methods. Comparing the results of the two methods, they show a specificity of 100% for all bacteria. The sensitivity for A. actinomycetemcomitans was 97.5%, for P. gingivalis 96.88%, and for P. intermedia 95.24%. The sensitivity for Tannerella forsythia and T. denticola was 100%. The Spearman correlation factor of two different measurements was 0.976 for A. actinomycetemcomitans, 0.967 for P. gingivalis, 0.949 for P. intermedia, 0.966 for Tannerella forsythia, and 0.917 for T. denticola. In conclusion, the in-house real-time PCR method developed in our laboratory can provide information about relative amount of five bacterial species present in subgingival plaque in patients with periodontitis. It is likely that such a test could be used in dental diagnostics in assessing the efficacy of any treatment to reduce the bacterial burden.

Microbiological Effects of Sodium Hypochlorite/-Amino Acids and Cross-linked Hyaluronic Acid Adjunctive to Non-surgical Periodontal Treatment.

PURPOSE: To investigate the microbiological outcomes obtained with either subgingival debridement (SD) in conjunction with a gel containing sodium hypochlorite and amino acids followed by subsequent application of a cross-linked hyaluronic acid gel (xHyA) gel, or with SD alone. MATERIALS AND METHODS: Forty-eight patients diagnosed with stages II-III (grades A/B) generalised periodontitis were randomly treated with either SD (control) or SD plus adjunctive sodium hypochlorite/amino acids and xHyA gel (test). Subgingival plaque samples were collected from the deepest site per quadrant in each patient at baseline and after 3 and 6 months. Pooled sample analysis was performed using a multiplex polymerase chain reaction (PCR)-based method for the identification of detection frequencies and changes in numbers of the following bacteria: Aggregatibacter actinomycetemcomitans (A.a), Porphyromonas gingivalis (P.g), Tannerella forsythia (T.f), Treponema denticola (T.d), and Prevotella intermedia (P.i). RESULTS: In terms of detection frequency, in the test group, statistically significant reductions were found for P.g, T.f, T.d and P.i (p < 0.05) after 6 months. In the control group, the detection frequencies of all investigated bacterial species at 6 months were comparable to the baseline values (p > 0.05). The comparison of the test and control groups revealed statistically significant differences in detection frequency for P.g (p = 0.034), T.d (p < 0.01) and P.i (p = 0.02) after 6 months, favouring the test group. Regarding reduction in detection frequency scores, at 6 months, statistically significant differences in favour of the test group were observed for all investigated bacterial species: A.a (p = 0.028), P.g (p = 0.028), T.f (p = 0.004), T.d (p <0.001), and P.i (p = 0.003). CONCLUSIONS: The present microbiological results, which are related to short-term outcomes up to 6 months post-treatment, support the adjunctive subgingival application of sodium hypochlorite/amino acids and xHyA to subgingival debridement in the treatment of periodontitis.

Multilocus Sequence Typing of Aggregatibacter actinomycetemcomitans Competently Depicts the Population Structure of the Species.

We developed a multilocus sequence typing scheme (MLST) for Aggregatibacter actinomycetemcomitans based on seven housekeeping genes, adk, atpG, frdB, mdh, pgi, recA, and zwf. A total of 188 strains of seven serotypes were separated into 57 sequence types. Whole-genome sequences were available for 140 strains, and in contrast to comparison of 16S rRNA genes, phylogenetic analysis of concatenated MLST gene fragments was in accordance with the population structure revealed by alignment of 785 core genes. MLST could not decisively identify the so-called JP2 clone associated with rapidly progressing periodontitis in adolescents, but noticeable clustering of JP2 genotype strains was revealed. The MLST scheme of A. actinomycetemcomitans can be assessed at www.pubmlst.org. IMPORTANCE Accurate diagnosis of infectious disease comprise identification, typing, and antimicrobial resistance of the infective agent. Bacteria are sometimes grouped within their species according to expression of specific toxins or particular antimicrobial resistance traits, but explicit typing for infection control and survey of pathogenesis necessitates genetic analysis such as multilocus sequence typing (MLST). Schemes for the most prevalent human pathogens have been available for more than 10 years, and time has come to extend the scrutiny to second-line infectious agents. One such pathogen is Aggregatibacter actinomycetemcomitans, which is commonly involved in periodontitis, and more rarely as the cause of infective endocarditis or spontaneous brain abscess. A MLST scheme for A. actinomycetemcomitans is now available at www.pubmlst.org. Whole-genome sequencing of a large number of isolates confirms that MLST competently depicts the population structure of the species.

Pulmonary Aggregatibacter actinomycetemcomitans infection masquerades as malignancy in a patient with periodontitis.

A 49-year-old man with a 37.5 pack-year smoking history presented with a suspected neoplasm of the right lung following the discovery of a metabolically active mass on positron emission tomography-CT imaging. The patient, who demonstrated poor oral hygiene, had a history of irregular problem-oriented dental visitation. Having excluded malignancy through histologic investigations, Aggregatibacter actinomycetemcomitans-a well-established periodontal pathogen-was subsequently cultured from his pulmonary aspirate. The patient was therefore managed with systemic antimicrobials and adjunctive dental extractions to eliminate the likely source of infection, whereafter the mass resolved without complication. This case corroborates previous reports of extraoral isolation of A. actinomycetemcomitans, which may mimic cancer clinically and radiographically. While a definitive causative link between untreated periodontitis and systemic infection remains to be elucidated, such cases present a compelling argument in favour of promoting oral health to prevent systemic disease.

The first study to detect co-infection of Entamoeba gingivalis and periodontitis-associated bacteria in dental patients in Taiwan.

Periodontitis, an inflammatory disease of the oral cavity, was caused by microbes from bacteria to protozoa. In this study, we detected protozoa, Entamoeba gingivalis and other three common pathogenic bacteria, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia by the Polymerase chain reaction (PCR) on patients.

Oral Lactobacillus strains reduce cytotoxicity and cytokine release from peripheral blood mononuclear cells exposed to Aggregatibacter actinomycetemcomitans subtypes in vitro.

BACKGROUND: This study evaluated the effect of oral lactobacilli on the cytotoxicity and cytokine release from peripheral blood mononuclear cells (PBMCs) when exposed to Aggregatibacter actinomycetemcomitans subtypes in vitro. The supernatants and cell wall extracts (CWEs) of eight A. actinomycetemcomitans strains, representing different subtypes, and three Lactobacillus strains were used. The PBMCs from six blood donors were exposed to supernatants and CWEs of A. actinomycetemcomitans or Lactobacillus strains alone or combinations and untreated cells as control. The cytotoxicity was determined by trypan blue exclusion method and IL-1ß secretion by ELISA. TNF-α, IL-6, and IL-8 secretions were measured using Bioplex Multiplex Immunoassay. RESULTS: Supernatants or CWEs from all bacterial strains showed cytotoxicity and IL-1ß secretion and the subtypes of A. actinomycetemcomitans showed generally a significantly higher effect on PBMCs than that of the Lactobacillus strains. Two highly toxic A. actinomycetemcomitans strains (JP2 and JP2-like) induced a higher response than all other strains. When combined, Lactobacillus significantly reduced the toxicity and the IL-1ß secretion induced by A. acinomycetemcomitans. The effect varied between the subtypes and the reduction was highest for the JP2 and JP2-like strains. The Lactobacillus paracasei strain SD1 had a higher reducing effect than the other Lactobacillus strains. This strain had a consistent reducing effect on all subtypes of A. actinomycetemcomitans cytotoxicity, and release of IL-1ß, IL-6, IL-8, and TNF-α from PBMCs of the blood donors. A strong and significant variation in cytokine release between the six blood donors was noticed. CONCLUSIONS: Lactobacillus spp. and L. paracasei SD1 in particular, showed a limited but statistically significant reducing interaction with A. actinomycetemcomitans toxicity and release of cytokines in vitro.

Aggregatibacter actinomycetemcomitans pneumonia mimicking lung cancer in a previously healthy 12-year-old child from Saudi Arabia: a case report.

Aggregatibacter actinomycetemcomitans formely known as Actinobacillus actinomycetemcomitans is a known part of the normal oral flora. It can sometimes cause oral or rarely extra-oral infections secondary to hematogenous extension or aspiration. It is a rare cause of invasive pneumonia. It can resemble tuberculosis or lung cancer in its presentation. Making the diagnosis in such case is crucial for better management that usually require tissue biopsy. We report a case of Aggregatibacter actinomycetemcomitans invasive pneumonia in a 12-year-old previously healthy boy from Saudi Arabia. After a prolonged course of antibiotics, clinical and radiological follow up showed complete resolution of the infection.

Effect of Periodontal Pathogens on Total Bone Volume Fraction: A Phenotypic Study.

Studies have shown that periodontal pathogens can enter the bloodstream, causing a series of reactions that can lead to a variety of systemic diseases. Epidemiological investigations also found a tight correlation between periodontitis (PD) and osteoporosis. This study aimed to further explore the effect of periodontal pathogens on bone volume fraction like bone tissue and mass, and explain the relationship between PD and osteoporosis. Sprague Dawley rats (female, 16 weeks old) were divided into the wild-type (WT) control group (n=9) and PD group (n=9). After eight weeks, periodontal tissues and ligatures, the fourth lumbar vertebra, the femur, the tibia, and blood were extracted and analyzed by micro-computed tomography (micro-CT), hematoxylin and eosin (H&E) staining, tartrate-resistant acid phosphatase (TRAP) staining, polymerase chain reaction (PCR), and enzyme-linked immunoassay (ELISA), respectively. We found that the bone mass of the lumbar vertebra, femur, and tibia was decreased in the PD group. The number of osteoclasts was higher in bone tissue in the PD group than in the WT group (P<0.05). The levels of inflammatory mediators and type I collagen C-terminal peptide (CTX-1) were higher in the PD group than in the WT group (P<0.05), although no significant difference in bone glutamic acid protein (BGP) levels was observed (P>0.05). In addition, we detected several periodontal pathogens, such as Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, and Fusobacterium nucleatum, in blood samples from rats in the PD group. These findings suggest that periodontal pathogens can enter the blood circulation from periodontal tissue, promote a systemic inflammation response, and subsequently reduce systemic bone density.

Aggregatibacter actinomycetemcomitans serotypes and JP2 outcomes related to clinical status over 6 years under periodontal maintenance therapy.

OBJECTIVE: This 6-year study evaluatedAggregatibacter actinomycetemcomitans outcomes and their relationship to clinical status. DESIGN: From the eligible individuals (23-70 years of age), 31 regular compliers (between-visit interval < 6 months) were randomly selected and matched for age/sex with 31 irregular compliers (between-visit interval > 6 months). Periodontal clinical examination and subgingival samples were obtained 5 times: T1 (baseline), T2 (after active periodontal therapy), T3 (2 years), T4 (4 years), and T5 (6 years). Total bacteria load, A. actinomycetemcomitans, and red complex species Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola levels were determined by qPCR while PCR was used to determine the occurrence of the a-b-c-d-e-f-g serotypes and the JP2 clone of A. actinomycetemcomitans. Data between groups was compared over time. RESULTS: At baseline PCR revealed A. actinomycetemcomitans prevalence of 9.7 % and JP2 prevalence of 6.7 %. A. actinomycetemcomitans qPCR levels were higher among individuals < 35 years of age and increased at T2 in irregular compliers. At in irregular compliers at the three follow-up visits. Serotypes a, d, and f showed greater values in at least one follow-up visit in regular compliers. A. actinomycetemcomitans showed negative correlation with probing depth (PD) while serotype b showed negative correlations with PD, PI, clinical attachment level and red complex. CONCLUSIONS: Longitudinally, compliance during PMT contributed to lower A. actinomycetemcomitans levels with some degree of correlation with clinical status. However, this study failed to report any positive effect on the occurrence of the most virulent representatives, i.e. serotype b and the JP2 clone.

Clinical and microbiological evaluation of non-surgical periodontal therapy in obese and non-obese individuals with periodontitis: a 9-month prospective longitudinal study.

Objective Obesity is a chronic disease that negatively affects an individual's general and oral health. The present study aimed to compare the clinical and microbiological effects of non-surgical periodontal therapy with the full mouth disinfection (FMD) protocol on obese and non-obese individuals at 9 months post-therapy. Methodology This clinical study was first submitted and approved by the Ethics Committee. Fifty-five obese patients and 39 non-obese patients with periodontitis were evaluated. The full-mouth periodontal clinical parameters, clinical attachment level (CAL), probing depth (PD), gingival index (GI), and plaque index (PI), were monitored at baseline, 3, 6, and 9 months after periodontal treatment with full mouth disinfection (FMD) protocol. The mean count of Tannerella forsythia , Porphyromonas gingivalis , Treponema Denticola , and Aggregatibacter actinomycetemcomitans was determined by quantitative polymerase chain reaction on subgingival biofilm samples. Demographic data were assessed by Chi-square test. For clinical and microbiological parameters, two-factor repeated-measures ANOVA was used. Results In both groups, periodontal therapy using the one-stage full-mouth disinfection protocol significantly improved CAL, PD, GI, and PI (p<0.05). Obese and non-obese patients equally responded to non-surgical periodontal therapy (p>0.05). Microbial count found no major differences (p>0.05) between obese and non-obese individuals who had undergone non-surgical periodontal therapy. Conclusions Obesity did not affect the clinical and microbiological outcomes of non-surgical periodontal therapy.

Er,Cr:YSGG laser surface treatment of gamma titanium aluminide: Scanning electron microscopy-energy-dispersive X-ray spectrometer analysis, wettability and Eikenella corrodens and Aggregatibacter actinomycetemcomitans bacteria count-in vitro study.

Dental implants play an important role in oral health. Titanium dental implants must endure the complex microflora environment of the oral cavity. Moreover, bacterial infections have been considered as one of the most important factors of implant failure. The issue of dental improvement through modification of chemical composition and surface treatment has received considerable critical attention. γ-TiAl as a novo biocompatible material revealed a slower corrosion rate in biological media rather Ti-6Al-4V. The objective of this study is to investigate the effect of Er,Cr:YSGG laser on γ-TiAl in comparison with sandblasted and acid-etched samples as the control groups and machined samples.Wettability, surface roughness, surface topography, scanning electron microscopy-energy dispersive X-ray spectrometer analysis of surface and subsurface of samples were investigated and bacteria counts of two periodontal bacterial strains (Aggregatibacter actinomycetemcomitans and Eikenella corrodens) were evaluated on the Er,Cr:YSGG laser surface-treated sandblasted and acid-etched and machined samples.The results of this investigation show that Er,Cr:YSGG laser surface treatment affects surface roughness, surface topography, wettability, chemical composition of the surface and bacteria count. Scanning electron microscopy-energy dispersive X-ray spectrometer analysis of the sample revealed the increment of titanium and oxygen content and reduction of aluminum content in the surface and subsurface layer. A. actinomycetemcomitans and E. corrodens count were found from the lowest level to highest in the sandblasted and acid-etched samples, laser samples and machined samples, respectively.Using controlled parameters of Er,Cr:YSGG laser ensured no significant adverse alteration. The findings to emerge from this study revealed the significant correlation between microbial count and wettability. Furthermore, the contact angle strongly correlated with surface roughness.

The effect of periodontal bacteria infection on incidence and prognosis of cancer: A systematic review and meta-analysis.

BACKGROUND: Periodontal bacteria is the major pathogens in the oral cavity and the main cause of adult chronic periodontitis, but their association with incidence and prognosis in cancer is controversial. The aim of this study was to evaluate the effect of periodontal bacteria infection on incidence and prognosis of cancer. METHODS: A systematic literature search of PubMed, Embase, Web of Science, and Cochrane Library databases was performed to obtain 39 studies comprising 7184 participants. The incidence of cancer was evaluated as odd ratios (OR) with a 95% confidence interval (95% CI) using Review Manager 5.2 software. Overall survival, cancer-specific survival and disease-free survival, which were measured as hazard ratios (HR) with a 95% CI using Review Manager 5.2 software. RESULTS: Our results indicated that periodontal bacteria infection increased the incidence of cancer (OR = 1.25; 95%CI: 1.03-1.52) and was associated with poor overall survival (HR = 1.75; 95% CI: 1.40-2.20), disease-free survival (HR = 2.18; 95%CI: 1.24-3.84) and cancer-specific survival (HR = 1.85, 95%CI: 1.44-2.39). Subgroup analysis indicted that the risk of cancer was associated with Porphyromonas gingivalis (Pg) infection (OR = 2.16; 95%CI: 1.34-3.47) and Prevotella intermedia (Pi) infection (OR = 1.28; 95%CI: 1.01-1.63) but not Tannerella forsythia (Tf) (OR = 1.06; 95%CI: 0.8-1.41), Treponema denticola (Td) (OR = 1.30; 95%CI: 0.99-1.72), Aggregatibacter actinomycetemcomitans (Aa) (OR = 1.00; 95%CI: 0.48-2.08) and Fusobacterium nucleatum (Fn) (OR = 0.61; 95%CI: 0.32-1.16). CONCLUSION: This meta-analysis revealed periodontal bacteria infection increased the incidence of cancer and predicted poor prognosis of cancer.

Hand Cellulitis and Abscess From a Kinkajou Bite: a Case Report and Review of Kinkajou Bites in Humans.

Medical literature on the microbiology of kinkajou bites is scarce. We present a patient presenting with hand cellulitis and abscess after a kinkajou bite that grew Aggregatibacter actinomycetemcomitans and Fusobacterium nucleatum. We review the literature regarding microbiology and management of kinkajou bites.

Clinical and microbiological evaluation of non-surgical periodontal therapy in obese and non-obese individuals with periodontitis: a 9-month prospective longitudinal study

Abstract Objective Obesity is a chronic disease that negatively affects an individual's general and oral health. The present study aimed to compare the clinical and microbiological effects of non-surgical periodontal therapy with the full mouth disinfection (FMD) protocol on obese and non-obese individuals at 9 months post-therapy. Methodology This clinical study was first submitted and approved by the Ethics Committee. Fifty-five obese patients and 39 non-obese patients with periodontitis were evaluated. The full-mouth periodontal clinical parameters, clinical attachment level (CAL), probing depth (PD), gingival index (GI), and plaque index (PI), were monitored at baseline, 3, 6, and 9 months after periodontal treatment with full mouth disinfection (FMD) protocol. The mean count of Tannerella forsythia , Porphyromonas gingivalis , Treponema Denticola , and Aggregatibacter actinomycetemcomitans was determined by quantitative polymerase chain reaction on subgingival biofilm samples. Demographic data were assessed by Chi-square test. For clinical and microbiological parameters, two-factor repeated-measures ANOVA was used. Results In both groups, periodontal therapy using the one-stage full-mouth disinfection protocol significantly improved CAL, PD, GI, and PI (p<0.05). Obese and non-obese patients equally responded to non-surgical periodontal therapy (p>0.05). Microbial count found no major differences (p>0.05) between obese and non-obese individuals who had undergone non-surgical periodontal therapy. Conclusions Obesity did not affect the clinical and microbiological outcomes of non-surgical periodontal therapy.

Microbiological study of the subgingival biofilm in HIV+/HAART patients at a specialized dental service / Estudio microbiológico del biofilm subgingival de pacientes VIH+ bajo TARGA en un servicio dental especializado

ABSTRACT The aim of this study was to describe the microbiological profile of HIV patients under highly active antiretroviral treatment (HAART). This crosssectional study comprised 32 HIV patients with periodontal disease (PD) who had been under HAART for more than 6 months. Information about the patients' medical history was obtained from clinical records. Clinical dental examination was performed by a calibrated researcher using standard dental instruments to determine probing depth (PD), clinical attachment level (CAL), and bleeding on probing (BOP). A total 4,765 periodontal sites were evaluated, 125 of which were also studied microbiologically. Subgingival biofilm samples were obtained using sterile paper points; one set was used for microbiological culture studies and the other for endpoint PCR. Statistical analysis was performed using KruskalWallis and posthoc DunnBonferroni contrast tests. All participants were on HAART at the time of the study, and 90.6% had a viral load below 50 copies/mm3. Prevalence of periodontally active sites was low in the study population. Microbiological studies: Black pigmented anaerobic bacteria and fusiform CFU counts were significantly higher in samples from sites with BOP and PD ≥4mm (p 0.020 and p 0.005, respectively). Molecular Assays: Detection of Porphyromonas gingivalis (p 0.002), Tannerella forsythia (p 0.023) and Treponema denticola (p 0.015) was significantly more frequent at sites with BOP and PD ≥4mm. Conclusions: The patients living with HIV/AIDS under HAART studied here had low prevalence of clinical periodontal disease signs. However, significant detection of P. gingivalis, T. denticola, and T. forsythia in periodontal active sites, and the involvement of these microorganisms as potential HIV reactivators, show the importance of creating awareness among dental health professionals of the need for close dental and periodontal monitoring in HIV patients.

RESUMEN El objetivo de este estudio fue describir el perfil microbiológico del biofilm subgingival de los pacientes con VIH bajo tratamiento antirretroviral de alta actividad (TARGA). El estudio comprendió a 32 pacientes VIH seropositivos con enfermedad periodontal (EP) que se encontraran en tratamiento con TARGA por más de 6 meses. Los antecedentes médicos de los pacientes se obtuvieron de las historias clínicas. El examen clínico instrumental (profun didad de sondaje (PS), nivel de inserción clínico (NIC) y sangrado al sondaje (SS)) fue realizado con instrumental odontológico estándar por un investigador calibrado. De este modo, se evaluaron un total de 4.765 sitios periodontales de los cuales 125 fueron estudiados microbiológicamente. Las muestras de biope lícula subgingival se obtuvieron empleando conos de papel estéril. Las muestras se emplearon en estudios microbiológicos y moleculares por PCR de punto final. El análisis estadístico se realizó según KruskalWallis y pruebas de contrastes posthoc de DunnBonferroni. El 90,6% de la población en estudio presentó carga viral inferior a 50 copias/mm3. La prevalencia de sitios periodontales activos fue baja (1%). Los recuentos de bacterias anaerobias estrictas pigmentadas de negro y fusiformes fueron significativamente más altos en muestras de sitios periodontales con SS positivo y PS ≥4 mm (p 0.020 y p 0.005). La detección molecular de Porphyromonas gingivalis (p 0.002), Tannerella forsythia (p 0.023) y Treponema denticola (p 0.015) fue significativamente mayor en los sitios con SS y PS ≥4mm. La prevalencia del 1% de enfermedad periodontal en el grupo de pacientes estudiados fue menor a la esperada, sin embargo; la detección significativa de P. gingivalis, T. denticola y T. forsythia en sitios periodontales activos y su potencial participación como agentes reactivadores del VIH, nos alerta de la importancia de crear conciencia en los profesionales de la salud (médicos y odontólogos) acerca de la necesidad de un monitoreo minucioso del estado periodontal de pacientes con características semejantes a las descriptas en la muestra poblacional estudiada.

Identification of Periopathogenes from Dental Plaque in Periodontal Patients with PCR Technique and Their Association with Composite Interleukin-1 Genotype.

INTRODUCTION: The present study aimed to assess the presence of main types of microorganisms involved in the aetiopathogenesis of chronic periodontitis with PCR technique and determinates the presence of composite IL-1 genotype and their associations with founded bacteria. MATERIAL AND METHOD: The examined group was consisted from 20 subjects with diagnosed chronic periodontitis and 20 healthy control without periodontitis. Clinical parameters like gingival index (GI), plaque index (PI), bleeding on probing (BOP), periodontal pocket depth (PPD) and clinical attachment lost (CAL) were determinates. Subgingival dental plaque was collected using a sterilized paper point. We used Parodontose Plus test, reverse hybridization kit, for the detection of periodontal marker bacteria, as well as for the detection of composite Interleukin -1 Genotype Results: The most present bacterial species detected from subgingival dental plaque was Treponema denticola and Porfiromonas gingivalis which was present in 65% of examined patients. In relation to the presence of positive genotype in patients, there was no significant difference between the test and control group for p> 0.05 (p = 1.00). For χ2=8,17 (p=0,06, p<0,05) there is an association between Prevotella intermedia, and composite genotype. Between positive genotype and analyzed bacterial species A. actinomycetem comitans for p> 0.05 (p = 1.00), P. gingivalis for p> 0.05 (p = 0.16), T. Forsythia for p> 0.05 (p = 0.20), T. Denticola for p> 0.05 (p = 0.64) no association was found. CONCLUSION: This investigations confirmed the strong association of these five examined periopathogenes with periodontitis.

Subgingival microbiological profile of periodontitis patients in Dominican Republic.

Several studies have tried to associate the presence of different pathogens with the onset and progression ofperiodontitis, reporting a wide variety of results from different populations and environments. The aim of this study was to determine the main periodontal pathogens present in the subgingival biofilm of Dominican patients with periodontitis, by using specific microbiological culturing techniques. Periodontitis patients were selected after a full-mouth periodontal evaluation, and assigned to different periodontitis groups based on percentage of affected locations. Subgingival samples were collected and analyzed by means of specific culture techniques. Anaerobic counts, frequency of detection and proportions of target pathogens were calculated. Variables were analyzed by means of Student's T-test or chi-square test. Twenty-nine subjects were recruited, of whom 17 were diagnosed with generalized periodontitis (GenP) and 12 with localized periodontitis (LocP). The most prevalent bacterial species in both groups was Prevotella intermedia (94.1% in GenP and 91.7% in LocP), followed by Porphyromonas gingivalis (88.2% in GenP and 83.3% in LocP). Total microbiota in subgingival samples was 1.3 x107 colony-forming units (CFU)/mL (standard deviation, SD=1.5 x107) and 9.6x10s CFU/mL (SD=1.1 x107) in GenP and LocP subjects, respectively, though differences were not statistically significant (p=0.222). The highest counts were observed for P gingivalis in both groups, with mean concentration 2.5x10s CFU/mL (6.1x10s) in GenP and 2.9x10s CFU/mL (5x10s) in LocP, with no statistically significant difference (p=0.879). These results suggest that relevant periodontal pathogens are found with diversity and abundance in the subgingival microbiota of adult Dominican patients with periodontitis.

Varios estudios han tratado de asociar la presencia de diferentes patógenos con el inicio y la progresión de la periodontitis, mostrando una gran variedad de resultados en diferentes poblaciones y entornos. El objetivo del presente estudio fue determinar los principales patógenos periodontales presentes en la biopelícula subgingival de pacientes dominicanos con periodontitis, utilizando técnicas específicas de cultivo microbiológico. Los pacientes con periodontitis se seleccionaron después de una evaluación periodontal de boca completa y se asignaron a diferentes grupos de periodontitis según el porcentaje de localizaciones afectadas. Las muestras subgingivales fueron recolectadas y analizadas mediante técnicas de cultivo específicas. Se calcularon los recuentos anaeróbicos, la frecuencia de detección y las proporciones de los patógenos seleccionados. Las variables se analizaron mediante la prueba T de Student o la prueba de chi-cuadrado. Se reclutaron veintinueve sujetos, 17 diagnosticados como periodontitis generalizada (GenP) 12 con periodontitis localizada (LocP). La especie bacteriana más prevalente en ambos grupos fue Prevotella intermedia (94.1% y 91.7%, respectivamente) y seguida de Porphyromonas gingivalis (88.2% y 83.3%, respectivamente). La microbiota total en muestras subgingivales fue 1.3 x107 unidades formadoras de colonias (CFU)/mL (desviación estándar, SD=1.5 x107) y 9.6x106 CFU / mL (SD=1.1 x107) en sujetos GenP y LocP, respectivamente, pero no hubo diferencias estadísticamente significativas (p=0.222). Los recuentos más altos se observaron para P. gingivalis en ambos grupos, con una concentración media de 2.5x106 CFU/mL (6.1x106) en GenP y 2.9x106 CFU/mL (5x106) en LocP, sin diferencias estadísticamente significativas (p=0.879). Estos resultados sugieren que se encuentran patógenos periodontales relevantes con diversidad y abundancia en la microbiota subgingival de pacientes adultos dominicanos con periodontitis.

Oral Bacterial Signatures in Cerebral Thrombi of Patients With Acute Ischemic Stroke Treated With Thrombectomy.

Background Chronic infections have been reported to be risk factors for both coronary heart disease and ischemic stroke. DNA of oral bacteria, mainly from the viridans streptococci group, has been detected in coronary thrombus aspirates of myocardial infarction and cerebral aneurysms. Viridans streptococci are known to cause infective endocarditis and possess thrombogenic properties. We studied the presence of oral bacterial DNA in thrombus aspirates of patients with acute ischemic stroke treated with mechanical thrombectomy. Methods and Results Thrombus aspirates and arterial blood were taken from 75 patients (69% men; mean age, 67 years) with acute ischemic stroke. The presence of Streptococcus species, mainly the Streptococcus mitis group, belonging to viridans streptococci as well as Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans in samples were determined using a quantitative polymerase chain reaction with specific primers and probes. The relative amount of bacterial DNA in a sample was determined with the comparative threshold cycle method. Bacterial DNA was detected in 84% (n=63) of aspired thrombi, and 16% (n=12) of samples were considered bacterial DNA negative. DNA of Streptococcus species, mainly the S mitis group, was found in 79% (n=59) of samples. The median relative amount of Streptococcus species DNA was 5.10-fold higher compared with the control blood samples from the same patients. All thrombi were negative for both P gingivalis and A actinomycetemcomitans. Conclusions This is the first study showing the common presence of bacterial DNA from viridans streptococci in aspired thrombi of patients with acute ischemic stroke. Streptococcal bacteria, mostly of oral origin, may contribute to the progression and thrombotic events of cerebrovascular diseases.

Microbiological study of the subgingival biofilm in HIV+/HAART patients at a specialized dental service.

The aim of this study was to describe the microbiological profile of HIV patients under highly active antiretroviral treatment (HAART). This crosssectional study comprised 32 HIV patients with periodontal disease (PD) who had been under HAART for more than 6 months. Information about the patients' medical history was obtained from clinical records. Clinical dental examination was performed by a calibrated researcher using standard dental instruments to determine probing depth (PD), clinical attachment level (CAL), and bleeding on probing (BOP). A total 4,765 periodontal sites were evaluated, 125 of which were also studied microbiologically. Subgingival biofilm samples were obtained using sterile paper points; one set was used for microbiological culture studies and the other for endpoint PCR. Statistical analysis was performed using KruskalWallis and posthoc DunnBonferroni contrast tests. All participants were on HAART at the time of the study, and 90.6% had a viral load below 50 copies / mm3. Prevalence of periodontally active sites was low in the study population. Microbiological studies: Black pigmented anaerobic bacteria and fusiform CFU counts were significantly higher in samples from sites with BOP and PD ≥4mm (p 0.020 and p 0.005, respectively). Molecular Assays: Detection of Porphyromonas gingivalis (p 0.002), Tannerella forsythia (p 0.023) and Treponema denticola (p 0.015) was significantly more frequent at sites with BOP and PD ≥4mm. Conclusions: The patients living with HIV/AIDS under HAART studied here had low prevalence of clinical periodontal disease signs. However, significant detection of P. gingivalis, T. denticola, and T. forsythia in periodontal active sites, and the involvement of these microorganisms as potential HIV reactivators, show the importance of creating awareness among dental health professionals of the need for close dental and periodontal monitoring in HIV patients.

El objetivo de este estudio fue describir el perfil microbiológico del biofilm subgingival de los pacientes con VIH bajo tratamiento antirretroviral de alta actividad (TARGA). El estudio comprendió a 32 pacientes VIH seropositivos con enfermedad periodontal (EP) que se encontraran en tratamiento con TARGA por más de 6 meses. Los antecedentes médicos de los pacientes se obtuvieron de las historias clínicas. El examen clínico instrumental (profun didad de sondaje (PS), nivel de inserción clínico (NIC) y sangrado al sondaje (SS)) fue realizado con instrumental odontológico estándar por un investigador calibrado. De este modo, se evaluaron un total de 4.765 sitios periodontales de los cuales 125 fueron estudiados microbiológicamente. Las muestras de biope lícula subgingival se obtuvieron empleando conos de papel estéril. Las muestras se emplearon en estudios microbiológicos y moleculares por PCR de punto final. El análisis estadístico se realizó según KruskalWallis y pruebas de contrastes posthoc de DunnBonferroni. El 90,6% de la población en estudio presentó carga viral inferior a 50 copias/mm3. La prevalencia de sitios periodontales activos fue baja (1%). Los recuentos de bacterias anaerobias estrictas pigmentadas de negro y fusiformes fueron significativamente más altos en muestras de sitios periodontales con SS positivo y PS ≥4 mm (p 0.020 y p 0.005). La detección molecular de Porphyromonas gingivalis (p 0.002), Tannerella forsythia (p 0.023) y Treponema denticola (p 0.015) fue significativamente mayor en los sitios con SS y PS ≥4mm. La prevalencia del 1% de enfermedad periodontal en el grupo de pacientes estudiados fue menor a la esperada, sin embargo; la detección significativa de P. gingivalis, T. denticola y T. forsythia en sitios periodontales activos y su potencial participación como agentes reactivadores del VIH, nos alerta de la importancia de crear conciencia en los profesionales de la salud (médicos y odontólogos) acerca de la necesidad de un monitoreo minucioso del estado periodontal de pacientes con características semejantes a las descriptas en la muestra poblacional estudiada.