Oral human cytomegalovirus prevalence and its relationships with periodontitis and Porphyromonas gingivalis in Japanese adults: a cross-sectional study

Abstract Objective This study aimed to clarify the association between oral human cytomegalovirus (HCMV) and periodontitis in Japanese adults. Methodology In total, 190 patients (75 men and 115 women; mean age, 70.2 years) who visited Hiroshima University Hospital between March 2018 and May 2020 were included. Oral rinse samples were taken to examine the presence of HCMV DNA using real-time polymerase chain reaction (PCR). P. gingivalis was detected by semi-quantitative PCR analysis. Results HCMV DNA was present in nine of 190 patients (4.7%). There were significant associations between HCMV presence and the presence of ≥4-mm-deep periodontal pockets with bleeding on probing (BOP) (P<0.01) and ≥6-mm-deep periodontal pockets with BOP (P=0.01). However, no significant relationship was observed between HCMV presence and periodontal epithelial surface area scores. Logistic regression analysis revealed that the presence of ≥4-mm-deep periodontal pockets with BOP was significantly associated with HCMV (odds ratio, 14.4; P=0.01). Propensity score matching was performed between patients presenting ≥4-mm-deep periodontal pockets with BOP (i.e., active periodontitis) and patients without ≥4-mm-deep periodontal pockets with BOP; 62 matched pairs were generated. Patients who had ≥4-mm-deep periodontal pockets with BOP showed a higher rate of HCMV presence (9.7%) than those who lacked ≥4-mm-deep periodontal pockets with BOP (0.0%). There was a significant relationship between HCMV presence and ≥4-mm-deep periodontal pockets with BOP (P=0.03). A significant relationship was found between HCMV/P. gingivalis DNA presence and ≥4-mm-deep periodontal pockets with BOP (P=0.03). Conclusions Coinfection of oral HCMV and P. gingivalis was significantly associated with active periodontitis. Moreover, interactions between oral HCMV and P. gingivalis may be related to the severity of periodontal disease.

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.

Investigations of the prevalence and virulence of Candida albicans in periodontal and endodontic lesions in diabetic and normoglycemic patients

Abstract Pulpal and periodontal tissues have similar microbiota that allows cross-contamination between the pulp and periodontal tissues. Objective The aim of this study was to investigate the prevalence of isolated Candida albicans from periodontal endodontic lesions in diabetic and normoglycemic patients, and the fungi's virulence in different atmospheric conditions. Material and Methods A case-control study was conducted on 15 patients with type 2 diabetes mellitus (G1) and 15 non-diabetics (G2) with periodontal endodontic lesions. Samples of root canals and periodontal pockets were plated on CHROMagar for later identification by polymerase chain reaction (PCR) and virulence test. Results C. albicans was identified in 79.2% and 20.8% of the 60 samples collected from diabetic and normoglycemic patients, respectively. Of the 30 samples collected from periodontal pockets, 13 showed a positive culture for C. albicans, with 77% belonging to G1 and 23% to G2. Of the 11 positive samples from root canals, 82% were from G1 and 18% from G2. Production of proteinase presented a precipitation zone Pz<0.63 of 100% in G1 and 72% in G2, in redox and negative (Pz=1), under anaerobic conditions in both groups. Hydrophobicity of the strains from G1 indicated 16.4% with low, 19.3% with moderate, and 64.3% with high hydrophobicity in redox. In G2, 42.2% had low, 39.8% had moderate, 18% had high hydrophobicity in redox. In anaerobic conditions, G1 showed 15.2% with low, 12.8% with moderate, and 72% with high hydrophobicity; in G2, 33.6% had low, 28.8% had moderate, and 37.6% had high hydrophobicity. There was statistical difference in the number of positive cultures between G1 and G2 (p<0.05) with predominance in G1. There was statistical difference for all virulence factors, except hemolysis (p=0.001). Conclusions Candida albicans was isolated more frequently and had higher virulence in diabetic patients.

Relaciones endoperiodontales: una frontera transitable / Endoperiodontal relationships: a passable border

La existencia de una íntima conexión entre el conducto radicular y el ligamento periodontal, ha dado lugar a lo que se conoce como relaciones endoperiodontales. Esto se debe a la presencia de varias vías anatómicas de comunicación entre ambas entidades: el foramen apical, los forámenes laterales pertenecientes a conductos accesorios y conductillos dentinarios en zonas de ausencia del cemento dentario protector. Los microorganismos y sus agentes tóxicos tienen la capacidad de afectar ambos tejidos en esa interrelación física y biológica. El diagnóstico diferencial entre enfermedad endodóntica y periodontal es de vital importancia para la elección del tratamiento y el pronóstico a distancia.

The intimate connection between the root canal and theperiodontal ligament generates a concept that is known asendo-periodontal relationships. This is due to the presence ofseveral anatomical communications between them: apical foramen,lateral foramina and dentinal tubules denuded of theircementum coverage. Microorganisms and their toxic agentscan affect both tissues due to this physical and biological interrelation.Proper differential between endodontic and periodontaletiology is vital to the accurate choice of treatment andfor the long term prognosis.

Detection of cfxA2, cfxA3, and cfxA6 genes in beta-lactamase producing oral anaerobes

ABSTRACT Purpose The aim of this study was to identify β-lactamase-producing oral anaerobic bacteria and screen them for the presence of cfxA and BlaTEM genes that are responsible for β-lactamase production and resistance to β-lactam antibiotics. Material and Methods Periodontal pocket debris samples were collected from 48 patients with chronic periodontitis and anaerobically cultured on blood agar plates with and without β-lactam antibiotics. Presumptive β-lactamase-producing isolates were evaluated for definite β-lactamase production using the nitrocefin slide method and identified using the API Rapid 32A system. Antimicrobial susceptibility was performed using disc diffusion and microbroth dilution tests as described by CLSI Methods. Isolates were screened for the presence of the β-lactamase-TEM (BlaTEM) and β-lactamase-cfxA genes using Polymerase Chain Reaction (PCR). Amplified PCR products were sequenced and the cfxA gene was characterized using Genbank databases. Results Seventy five percent of patients carried two species of β-lactamase-producing anaerobic bacteria that comprised 9.4% of the total number of cultivable bacteria. Fifty one percent of β-lactamase-producing strains mainly Prevotella, Porphyromonas, and Bacteroides carried the cfxA gene, whereas none of them carried blaTEM. Further characterization of the cfxA gene showed that 76.7% of these strains carried the cfxA2 gene, 14% carried cfxA3, and 9.3% carried cfxA6. The cfxA6 gene was present in three Prevotella spp. and in one Porphyromonas spp. Strains containing cfxA genes (56%) were resistant to the β-lactam antibiotics. Conclusion This study indicates that there is a high prevalence of the cfxA gene in β-lactamase-producing anaerobic oral bacteria, which may lead to drug resistance and treatment failure.

Alteraciones morfo-cardiovasculares producidas por bacterias periodontopatógenas en modelo murino / Morphological and cardiovascular alterations produced by periodontopathic bacteria in a murine model

Diferentes autores han descrito a la enfermedad periodontal como un factor de riesgo de ateroesclerosis. Este estudio tuvo como objetivo determinar la posible relación entre las bacterias periodontopatógenas y alteraciones cardiovasculares en modelos murinos con ingesta hiperlipídica. Se utilizaron 30 ratas Wistar divididas en un grupo control (n=5) y tres grupos experimentales (n=15): Grupo I (n=5): dieta normal e infectados; Grupo II (n=5): dieta hiperlipídica y sin infectar y Grupo III (n=5): dieta hiperlipídica e infectados. La infección del surco gingival se logró previa sedación a través de hilos de sutura contaminados, colocados durante 6 días consecutivos, con Pophyromonas gingivalis (P. gingivalis) (5x10a-4 UFC) y Agregatibacter actinomycetemcomitans (A. actinomycetemcomitans) (5x10-4 UFC) obtnidas de muestras de pacientes con periodontitis crónica. A los 30 días posinfección se fijaron muestras de corazón y arteria aorta para su procesamiento. En el grupo control no se encontraron alteraciones. En el grupo I se observó en el endocardio valvular abundante exudado fibrinoleucocitario y focos hemorrágicos. En el miocardio, neovascularización y sufusiones hemorrágicos y en la pared de vasos arteriales, engrosamiento intimal. En el grupo II se observaron depósitos lipídicos en el interior de las células endoteliales, estrías grasas y pequeños ateromas. El grupo III mostró en un 100 por ciento de los casos, placas ateromatosas bien detalladas con abundante depósito de colesterol, calcificaciones, hemorragias y neovascularización. La suma de estos factores de riesgo: dieta hiperlipídica e infección con bacterias periodontopatógenas, potenciarían la expresión de alteraciones cardíacas y coronarias, incrementando la formación de ateromas

Occurrence of Porphyromonas gingivalis and its antibacterial susceptibility to metronidazole and tetracycline in patients with chronic periodontitis

La periodontitis crónica es una enfermedad infecciosa multifactorialasociada a bacilos Gram-negativos anaeróbicos estrictos que se encuentran inmersos en la biopelícula subgingival. Porphyromonas gingivalis, importante patógeno periodontal, es frecuentemente detectado en pacientes con periodontitis crónica. Los aislamientos clínicos de P. gingivalis tienden a ser susceptibles a la mayoría de agentes antimicrobianos; sin embargo, se tiene poca información sobre la susceptibilidad antimicrobiana invitro. El objetivo de este estudio fue determinar la frecuencia de P. gingivalis en pacientes con periodontitis crónica y determinar la susceptibilidad antimicrobiana en términos de concentración inhibitoria mínima (CIM) de los aislamientos clínicos a metronidazol y tetraciclina. Se realizó un estudio observacional descriptivo enel que se incluyeron 87 pacientes con periodontitis crónica. Las muestras tomadas con conos de papel de la bolsa periodontal se depositaron en caldo tioglicolato, se incubaron durante 4 horas a 37 oC en anaerobiosis y se resembraron en agar anaeróbico Wilkins-Chalgren (Oxoid). La identitficación de los aislamientos serealizó con el sistema RapIDTM ANA II (Remel) y la susceptibilidadantibiótica para metronidazol y tetraciclina se evaluó mediante la técnica M.I.C.Evaluator (M.I.C.E., Oxoid). En 30 de los 87 pacientes con periodontitis crónica se identificó P. gingivalis, lo que representa una frecuencia de 34.5 por ciento. Todos los 30 aislamientos (100 por ciento) fueron sensibles al metronidazol con valores de CIM desde 0.015 hasta 4 ug/ml. En cuanto a tetraciclina, 27 aislamientos(90 por ciento) fueron sensibles con valores de CIM desde <0.015 hasta4 ug/ml; los restantes 3 aislamientos (10%) fueron resistentes a tetraciclina con valores de CIM de 8 ug/ml. En cuanto a edad, género, profundidad de bolsa, nivel de inserción clínico y severidad de la periodontitis no se presentaron diferencias estadísticamentesignificativas

Evaluación de la terapia mecánica periodontal en bolsas profundas: respuesta clínica y bacteriológica / Evaluation of periodontal mechanical therapy in deep pockets: Clinical and bacteriological response

Objetivos: Evaluar la composición microbiológica y los parámetros clínicos de bolsas periodontales >5 mm de profundidad al inicio, 1 semana, 3 y 12 meses post raspado y alisado radicular. Materiales y Métodos: Se tomaron registros clínicos y muestras de placa subgingival de 44 sitios de pacientes con diagnóstico de periodontitis crónica. Se identificaron por técnica de Reacción en Cadena de la Polimerasa (PCR) patógenos putativos periodontales: Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Treponema denticola (Td), Tannerella forsythia (Tf) y Prevotella intermedia (Pi). Los pacientes recibieron terapia mecánica periodontal y fueron reevaluados a los 7 días, 3 y 12 meses. Resultados: Luego del tratamiento, todos los parámetros clínicos (Placa Bacteriana, Hemorragia, Supuración, Profundidad al Sondaje y Nivel de Inserción Clínica) se redujeron significativamente y los valores obtenidos se mantuvieron hasta los 12 meses. Al inicio, las especies bacterianas prevalentes fueron Pg, presente en 66 por ciento de los sitios, Tf (55 por ciento) y Td (41 por ciento). Los sitios más profundos se relacionaron con las asociaciones Tf-Td (6.8 mm) y Tf-Td-Pi (7 mm). Post terapia, el número de sitios positivos para Td, Tf y Pg se redujo significativamente. Conclusiones: El raspado y alisado radicular mejoró significativamente los parámetros clínicos y redujo la prevalencia de los patógenos periodontales Pg, Tf y Td en bolsas periodontales profundas. Los resultados obtenidos se mantuvieron hasta los 12 meses. No se detectaron mayores pérdidas de inserción clínica en el 86 por ciento de los sitios a 3 meses y en 79 por ciento a los 12 meses. Los sitios en los que el tratamiento no fue efectivo en la eliminación de patógenos a los 12 meses desarrollaron mayores profundidades de sondaje.

Objectives: To evaluate the microbial composition and clinical parameters of periodontal pockets with probing depth >5 mm at baseline, 1 week, 3 and 12 months after scaling and root planning. Methods: Clinical parameters were measured and bacterial samples were collected from 44 sites in 11 patients with chronic periodontitis. By means of Polymerase Chain Reaction (PCR) the presence of Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Treponema denticola (Td), Tannerella forsythia (Tf) and Prevotella intermedia (Pi) was estimated. The patients received mechanical periodontal therapy and were evaluated after 1 week, 3 months and 12 months. Results: After treatment, all clinical parameters (Plaque, Bleeding on Probing, Supuration, Probing Pocket Depth and Clinical Attachment Level) were significantly reduced, and the values obtained were maintained up to the 12 months that the study lasts. At baseline, the most prevalent species were Pg, present in 66 percent of the sites, Tf (55 percent) and Td (41 percent). The deepest sites were related to the association Tf-Td (6.8 mm) and Tf-Td-Pi (7 mm). The number of positive sites for Td, Tf and Pg was significantly reduced after therapy. Conclusions: Scaling and root planning improve significantly clinical parameters as well as reduce the prevalence of periodontal pathogens Pg, Td and Tf in deep periodontal pockets. The results obtained were maintained up to 12 months. No further clinical attachment loss was found in 86 percent of the sites at 3 months and 79 percent at 12 months. The sites where the treatment failed in removing pathogens developed at 12 months greater probing pocket depths.

Efecto del tratamiento periodontal sobre la microbiota subgingival en pacientes con preeclampsia / Effect of periodontal therapy on the subgingival microbiota in preeclamptic patients

Introducción. Pocos estudios han descrito la microbiota subgingival en mujeres embarazadas con preeclampsia leve. Objetivo. Identificar cambios periodontales y de la microbiota subgingival en mujeres embarazadas con preeclampsia, después del tratamiento periodontal. Materiales y métodos. En un análisis secundario de un ensayo clínico de asignación aleatoria, se estudiaron 57 pacientes con preeclampsia en el Hospital Universitario del Valle de Cali. Se asignaron al azar 31 al grupo de intervención periodontal (detartraje y alisado subgingival ultrasónico y manual) durante su embarazo y otras 26 al grupo control (profilaxis supragingival). Se determinaron los parámetros clínicos periodontales y la microbiota subgingival a la inclusión al estudio y en el posparto. Se evaluaron 8 bacterias periodontopáticas y 2 virus herpes por reacción en cadena de la polimerasa. Se usaron las pruebas de ji al cuadrado, test de McNemar o t de Student, con un nivel de significancia de p≤ Resultados. Los grupos fueron comparables en las variables clínicas y microbiológicas al inicio del estudio. El tratamiento periodontal redujo el promedio de la profundidad de bolsa en el grupo de intervención de 2,44±0,31 a 2,31±0,24 mm (p=0,000) y en el grupo control de 2,58±0,37 a 2,44±0,39 mm (p=0,000),y el índice de sangrado, de 16,4±1,5 a 7,9±0,7 % en el primero (p=0,000), y de 17,1±1,8 a 10±0,9 %, en el segundo (p=0,002). La frecuencia de detección de microorganismos no varió de manera significativa entre los grupos. Conclusión. El raspaje y alisado radicular, así como la profilaxis supragingival, redujeron de manera significativa la profundidad a la sonda y el índice de sangrado gingival. El tratamiento periodontal no fue más efectivo que la profilaxis para reducir los organismos periodontopáticos o los virus herpes.

Introduction. Few studies have described subgingival microbiota in pregnant women with mild preeclampsia. Objective. Clinical periodontal and subgingival microbiota changes were identified in pregnant women with mild preeclampsia after periodontal treatment. Materials and methods. In a secondary analysis of a randomized clinical trial, 57 preeclamptic women were studied at Hospital Universitario del Valle in Cali, Colombia. Thirty one women were randomized to the periodontal intervention group (subgingival scaling and planing ultrasonic and manual) during pregnancy and 26 to the control group (supragingival prophylaxis). Periodontal clinical parameters and subgingival microbiota were characterized at the time of acceptance into the study and again at postpartum. Eight periodontopathic bacteria and 2 herpesviruses were assessed by polymerase chain reaction. Chi-square, McNemar or Student´s t tests were used, with a significance level of p≤0.05. Results. Both groups were comparable in the clinical and microbiological variables at baseline. Periodontal treatment reduced the average pocket depth in the intervention group from 2.4±0.3 to 2.3±0.2 mm (p<0.001) and in control group 2.6±0.4 to 2.44±0.4 mm, (p<0.001) and bleeding index 16.4±1.5% to 7.9±0.7% in the intervention group(p<0.001) and 17.1±1.8% to 10±0.9% in the control group (p=0.002). The frequency of detection of microorganisms did not differ significantly between groups. Conclusion. Scaling/root planning and supragingival prophylaxis significantly reduced the probing depth and gingival bleeding index. Periodontal treatment was not more effective than prophylaxis in reducing periodontopathic organisms or herpesvirus.

Honeybee propolis extract in periodontal treatment: A clinical and microbiological study of propolis in periodontal treatment.

Objective: This study was conducted to evaluate by clinical and microbiological parameters the effect of subgingival irrigation with propolis extract. Materials and Methods: Twenty patients diagnosed with chronic periodontitis, each presenting three non-adjacent teeth with deep pockets, were selected. Subgingival plaque sampling and clinical recording (at baseline) and scaling and root planing was performed. Two weeks later the selected periodontal sites were submitted to one of the following treatments: Irrigation with a hydroalcoholic solution of propolis extract twice a week for 2 weeks (group A); irrigation with a placebo twice a week for 2 weeks (group B); or no additional treatment (group C). Clinical and microbiological data was collected at baseline and after 4, 6, and 8 weeks. Results: A decrease in the total viable counts of anaerobic bacteria (P=.007), an increase in the proportion of sites with low levels (≤10 5 cfu/mL) of Porphyromonas gingivalis (P=.044), and an increase in the number of sites negative for bleeding on probing was observed in group A sites as compared to group B and C sites. Conclusion: Subgingival irrigation with propolis extract as an adjuvant to periodontal treatment was more effective than scaling and root planing as assessed by clinical and microbiological parameters.

Detection of odoriferous subgingival and tongue microbiota in diabetic and nondiabetic patients with oral malodor using polymerase chain reaction.

Background: Halitosis has been correlated with the concentration of volatile sulfur compounds (VSCs) produced in the oral cavity by metabolic activity of bacteria colonizing the periodontal pockets and the dorsum of the tongue. It has been assumed that there is a relationship between periodontal disease and diabetes mellitus. Objectives: The aim of the study was to assess the malodor using the organoleptic method and tanita device; to quantify odoriferous microorganisms of subgingival plaque and tongue coating, such as P. gingivalis (Pg), T. forsythia (Tf), and F. nucleatum (Fn) using polymerase chain reaction (PCR) in nondiabetic and diabetic chronic periodontitis patients. Patients and Methods: Thirty chronic periodontitis patients (with and without diabetes) with 5-7 mm pocket depth, radiographic evidence of bone loss, and presence of oral malodor participated in this study. Subjective assessment of mouth air was done organoleptically and by using a portable sulfide monitor. Tongue coating was also assessed. Results: The scores of plaque index, gingival index, gingival bleeding index, VSC levels, and tongue coating between the nondiabetic and diabetic patients were not significant (P>0.5). In nondiabetic patients, Fn was found to be significantly (P<0.5) more in tongue samples, whereas Pg and Tf have not shown significant values (P>0.5). In diabetic patients, Fn and Tf have shown significant (P<0.5) an increase in subgingival and tongue samples, respectively, whereas Pg has not shown significant difference between subgingival and tongue samples. Interpretation and Conclusion: The results confirm that there is no difference in clinical parameters between nondiabetic and diabetic periodontitis patients, but the odoriferous microbial profiles in tongue samples of diabetic patients were found to be high. However, there is a weak positive correlation between VSC levels, clinical parameters, and odoriferous microbial profiles.

Positive correlations between presence of Gram negative enteric rods and Porphyromonas gingivalis in subgingival plaque

La asociación entre bacilos entéricos Gram-negativos y Porphyromonas gingivalis en las enfermedades periodontales ha recibido poca atención en la literatura. Así, el objetivo de este estudio fue investigar la asociación entre bacilos entéricos Gram-negativos, Porphyromonas gingivalis y los parámetros clínicos de la periodontitis. Se evaluó la prevalencia de bacilos entéricos Gram-negativos y P. gingivalis en pacientes con periodontitis crónica. Se utilizaron pruebas de Chi-cuadrado y Mann-Whitney para determinar las diferencias en las variables clínicas frente a la presencia o ausencia de ambos microorganismos. Se empleó el coeficiente de correlación de Spearman para determinar las correlaciones de ambos organismos y los datos clínicos obtenidos. Se detectaron bacilos entéricos Gram-negativos y P. gingivalis en 20 (26,3 por ciento) y 51 (67,1 por ciento) sujetos, respectivamente. Un total de 17 (22,4 por ciento) individuos presentó los dos microorganismos estudiados. Se observó una correlación poisitiva significativa entre bacilos entéricos y la presencia de P. gingivalis (r=0,531m O<0,0001). Se encontró una correlación positiva estadísticamente significativa de los dos microorganismos estudiados con la profundidad de sondaje (PS), pérdida de inserción clínica y sangrado al sondaje (P<0,0001). El promedio de la profundidad de sondaje (mm), de los sitios incluidos en la muestra fue significativamente superior en pacientes con presencia de P. gingivalis y bacilos entéricos Gram-negativos. La presente investigación sugiere que la presencia de bacilos entéricos Gram negativos y P. gingivalis se relaciona con condiciones periodontales adversas. Estos resultados podrían tener un impacto en el tratamiento periodontal y se deben tener en cuenta en la terapia mecánica y antimicrobiana de la periodontitis en diferentes poblaciones.

Periodontal parameters and BANA test in patents with chronic renal failure undergoing hemodialysis

OBJECTIVES: The aim of this study was to analyze the periodontal parameters of patients with chronic renal failure. MATERIAL AND METHODS: The periodontal status of 16 Brazilian patients aged 29 to 53 (41.7±7.2) years with chronic renal failure (CRF) and another matched group of 14 healthy controls with periodontitis was assessed clinically and microbiologically. Probing pocket depth (PPD), gingival recession (GR), dental plaque index (PLI), gingival index (GI), and dental calculus index (CI) were the clinical parameters recorded for the entire dentition (at least 19 teeth), while the anaerobic periodontopathogen colonization in four sites with the highest PPD was evaluated using the BANA test ("PerioScan"; Oral B). RESULTS: The results for the CRF group and control group, respectively were: PPD: 1.77±0.32 and 2.65±0.53; GR: 0.58±0.56 and 0.51±0.36; PLI: 1.64±0.56 and 1.24±0.67; GI: 0.64±0.42 and 0.93±0.50; CI: 1.17±0.54 and 0.87±0.52. Comparison between groups using the "t" test revealed a significantly increased PPD (p<0.001) in the control group. Comparison of the other clincial parameters by the Mann-Whitney test showed differences only for PLI, which was significantly higher (p<0.05) in the CRF group. Spearman's test applied to each group showed a positive correlation among all clinical parameters, except for GR (p<0.05). None of the groups showed any correlation between GR and GI, while a significant negative correlation between GR and PPD was observed for the CRF group. The percentage of BANA-positive sites was 35.9 percent for the CRF group and 35.7 percent for the control group. The BANA test correlated positively with PPD only in the control group and with GR only in the CRF group. CONCLUSIONS: In spite of a higher PLI and dense anaerobic microbial population even in shallow PPD, patients with CRF exhibited better periodontal conditions than periodontitis patients, which is an evidence of altered response to local irritants.

Prevalence of Staphylococcus spp and Candida spp in the oral cavity and periodontal pockets of periodontal disease patients

The oral cavity can act as a reservoir of certain pathogens that can cause systemic infections. The periodontal pocket is an ecological niche appropriate for hosting microorganisms that could act as opportunistic pathogens. The ability of Staphylococcus spp and Candida spp to form a biofilm and live within certain niches allows them to develop mechanisms that increase persistence, such as the evasion of host defenses and antibiotic efficacy. These microorganisms can easily be or become resistant to antibiotics and lead to superinfection. The aims of this study were to assess the presence of Staphylococcus aureus and Staphylococcus spp in biofilm in subgingival plaque and oral cavity of individuals with gingival-periodontal disease, to identify isolates and the relationship with Candida spp. The study included eighty-two patients, aged 18-70 years with periodontal disease and at least two sites with probing depth ≥3 mm. Participants’ data were evaluated individually. Subgingival biofilm samples were obtained using Gracey curettes 7/8, after supragingival biofilm removal, and a sample from the oral cavity (buccal mucosa, tongue and cheek mucosa) by sterile swab. Of all the patients studied, 42.7% exhibited Staphylococcus spp in the periodontal pocket and 69.5% in the oral cavity while 25.6% exhibited Candida spp in the periodontal pocket and 42.7% in the oral cavity. However, 13.4% had both microorganisms in the periodontal pocket and 36.6% in the oral cavity. The prevalence of Staphylococcus aureus was 13.4% in the periodontal pocket and 15.8% in the oral cavity. Candida albicans was the most prevalent yeast in the periodontal pocket (76.2%) and in the oral cavity (63.0%).

La cavidad bucal puede actuar como reservorio de ciertos patógenos que pueden producir infecciones sistémicas. La bolsa periodontal es un nicho ecológico propicio para albergar microorganismos que podrian actuar como patógenos oportunistas. La posibilidad que Staphylococcus spp y Candida spp puedan formar un biofilm o biopelícula y vivir dentro de ciertos nichos les permite a estos microorganismos desarrollar ciertos mecanismos que aumentan su persistencia como ser la capacidad de eludir las defensas del huésped y la terapia antimicrobiana. Estos microorganismos pueden ser o fácilmente convertirse en resistentes a los antibióticos y dar origen a una supeinfección. El propósito de este estudio fue evaluar la presencia de Staphylococcus aureus y Staphylococcus spp en biofilm placa subgingival y en cavidad oral en sujetos con enfermedad gingivoperiodontal, identificar los microorganismos aislados y su relación con la portación de Candida spp. El estudio incluyo ochenta y dos pacientes, de edades entre 18 a 70 anos de edad, con enfermedad periodontal, y al menos dos sitios con la profundidad de sondaje ≥ 3 mm. Se evaluaron los datos individuales. Las muestras de biofilm subgingival fueron obtenidas con cureta tipo Gracey 7/8, previa remoción del biofilm supragingival y una muestra de cavidad oral (mucosa, lengua y carrillo) mediante hisopo estéril. Del total de los pacientes estudiados, el 42,7% mostraron Staphylococcus spp en la bolsa y el 69,5% en la cavidad oral, mientras que 25,6% mostraron Candida spp en la bolsa y 42,7% en la cavidad oral. Sin embargo, el 13,4% tenían ambos microorganismos en la bolsa y el 36,6% en la cavidad oral. La prevalencia de Staphylococcus aureus en la bolsa periodontal fue de 13,4% y 15,8% en la cavidad oral. Candida albicans fue la levadura más frecuente en la bolsa periodontal (76,2%) y en la cavidad oral (63,0%).

Expressão de fatores de virulência, mecanismos de resistência aos agentes antimicrobianos e análise molecular da resistência aos beta-lactâmicos de enterobactérias isoladas de bolsas periodontais / Expression of virulence factors, mechanisms of resistance to antimicrobial agents and molecular analysis of beta-lactam resistance to Enterobacteriaceae isolated from periodontal pockets

Em estudo anterior, as espécies de enterobactérias apresentando perfis variados de resistência aos antimicrobianos foram detectadas em 20% dos sítios com lesões periodontais de pacientes sadios do ponto de vista sistêmico. Tais cepas microbianas foram submetidas a investigações com o intuito de determinar à expressão de enzimas hidrolíticas para substratos diversos, a multirresistência aos agentes antimicrobianos e os mecanismos de resistência aos antimicrobianos da classe dos B lactâmicos. A maioria das amostras expressou atividade de gelatinase (65%), caseinase (30%) e elastase (10%). Lipase, lecitinase e DNase foram observadas apenas para Serratia marcescens. A multirresistência (considerado como a resistência a pelo menos dois agentes antimicrobianos de famílias diferentes) foi observada em 56% das amostras isoladas. A maioria das cepas foi resistente à ampicilina (93,75%) e amoxicilina/ácido clavulânico (81,25%). Investigações sobre a resistência aos antibióticos B-lactâmicos mostraram que três amostras resistentes à cefalosporinas de 2ª geração, apresentaram perfis plasmidiais de diferentes pesos moleculares. A expressão fenotípica de B-lacatamases, foi detectada nas cepas de Enterobacter cloacae (PcOM46 e PcOM5) e S. marcescens (PcOM63). No entanto, na análise molecular, não foi possível confirmar a expressão fenotípica de diferentes B-lactamases, com exceção do E. cloacae PcOM46, que apresentou amplificação para AmpC e blatem. Embora sensível à maioria dos antibióticos B-lactâmicos (exceção feita à ampicilina e amoxicilina/ácido clavulânico), amostra de S. marcescens PcOM68 apresentou amplificação para o gene blashv. Os experimentos de conjugação não detectaram a transferência de plasmídios para uma cepa de Escherichia coli K12 sensível aos B-lactâmicos, o mesmo ocorreu nos procedimentos de transformação por eletroporação e por CaCI2, sugerindo uma resistência dependente de genes cromossomiais. A expressão de diferentes atividades enzimáticas...

In a previous study, the enterobacterial species were detected in 20% of systemically healthy patients presenting periodontal lesions, with diferent profiles of antimicrobial resistance. These microbial strains were investigated in view to determine the expression of hydrolytic enzymes for diverse substrates, multiresistance to antimicrobial agents, and the mechanisms of resistance to beta-lactam antibiotics. The enzymes related to bacterial virulence were detected phenotypically in 90% isolates. The proteolytic activity displayed by the isolates against gelatin, casein and elastin was detected in 65%, 30% and 10%, respectively. Lipase, lecithinase, and DNase were observed only for Serratia marcescens strains. Multi-resistant phenotypes (considered as the resistance to at least two antimicrobial agents from different families) were observed for 56% enterobacterial isolates. Most of the strains were resistant to ampicillin (93.75%) and amoxicillin/clavulanic acid (81.25%). Investigations concerned to the resistance to beta-lactam antibiotics demonstrated that three bacterial strains resistant to penicilins and 2nd generation cephalosporins, had detectable plasmids. The extended resistance to B-lactams was detected phenotypically for E. cloacae (PcOM46 and PcOM5) and S. marcescens (PcOM63) strains. However, the molecular detection of extended spectrum beta-lactamase failed to confirm the phenotypic expression of different B-lactamases, with exception to the E. cloacae PcOM46 isolate, which presented amplification for both ampC and blatem. Although sensitive to most of the B-lactam antibiotics (exception made to ampicilin and ampicilin/clavulanate), the strain S. marcescens PcOM68 was shown to amplify the blashv gene. Plasmid transference by both conjugation and transformation procedures failed to detect the resistance by a B-lactam-sensitive strain (E. coli K12), suggesting a chromosomal dependent resistance. The expression of varied enzymatic activities...

Occurrence of Aggregatibacter actinomycetemcomitans in Brazilian indians from Umutina Reservation, Mato Grosso, Brazil

Aggregatibacter actinomycetemcomitans is associated with periodontal disease, especially localized aggressive periodontitis, produces a potent leukotoxin and its distribution is influenced by ethnic characteristics of the population. Objective: Using culture and polymerase chain reaction (PCR) techniques, this study evaluated the occurrence of this microorganism and the distribution of leukotoxic strains isolated from Indians belonging to the Umutima Reservation, Mato Grosso, Brazil. MATERIAL AND METHODS: Forty-eight native Brazilians with gingivitis and 38 with chronic periodontitis, belonging to Umutina, Paresi, Bororo, Bakairi, Kayabi, Irantxe, Nambikwara and Terena ethnicities, were studied. Subgingival, supragingival and saliva samples of each patient were collected and transferred to VMGA III medium and to ultra pure Milli Q water. Bacteria were grown on TSBV agar and incubated in anaerobiosis (90 percent N2 + 10 percent CO2) at 37ºC for 72 h. The presence of the ltx promoter was determined by PCR, and a 530 bp deletion in the promoter was evaluated by using specific primers. RESULTS: A. actinomycetemcomitans was isolated from 8.33 percent of saliva, supragingival and subgingival samples from patients with gingivitis and from 18.42 percent of saliva and supragingival biofilm, and 26.32 percent subgingival biofilm from patients with chronic periodontitis. By PCR, the bacterial DNA was detected in 8.33 percent of saliva, supragingival and subgingival biofilms from patients with gingivitis and from 23.68 percent of saliva, 28.95 percent supragingival biofilm and 34.21 percent subgingival biofilm from patients with periodontitis. All strains were grouped as non-JP2 clones based on the absence of deletion in the leukotoxin promoter. Differences among the microbial and clinical parameters in patients were analyzed by using the Mann-Whitney, Chi-square or Fisher's exact tests. CONCLUSIONS: The present results suggest that A. actinomycetemcomitans ...

Community periodontal index of treatment needs index: An indicator of anaerobic periodontal infection.

Background : Community periodontal index of treatment needs (CPITN) index is commonly used to measure periodontal disease. It's uniqueness, apart from assessing the periodontal status, also gives the treatment needs for the underlying condition. Benzoyl-DL-arginine napthylamide (BANA) test is a chair side diagnostic test used to detect the presence of putative periodontal pathogens. We correlated the CPITN scores of patients with BANA test results to assess the validity of CPITN as an indicator of anaerobic periodontal infection. Objectives : The present study was aimed to correlate the CPITN scores with the BANA activity of subgingival plaque. The objective was to assess the validity of CPITN index as indicator of anaerobic periodontal infection. Patients and Methods : A total of 80 sites were selected from 20 patients with generalized chronic periodontitis. After measuring the probing depth with CPITN C probe, the highest score from each sextant was selected according to the CPITN criteria and subgingival plaque samples were collected using a sterile curette and the BANA test was performed. Results : Kendall's tau-b and Chi- square test were used to assess the correlation between the BANA test results and CPITN scores. Results indicated sensitivity (92.86%), specificity (80%) and agreement (91.25%); indicating the validity of CPITN in assessing anaerobic infection. Conclusion : There was a significant correlation between BANA test results and scores 3 and score 4 of CPITN index (P < 0.001) clearly indicating the presence of anaerobic periodontal infection.

Enhanced susceptibility of Candida albicans to chlorhexidine under anoxia

Aim: Periodontal pockets can be colonized not only by bacteria, but also by Candida albicans. However, its role in periodontitis is unknown. This study evaluated the inhibitory performance of chlorhexidine digluconate under normoxic and anoxic conditions against 16 strains of C. albicans from periodontal pockets and other 20 from the oral mucosa. Methods: Strains were grown in normoxia and anoxia to adapt themselves to the different atmospheric conditions. Microdilution-based assays were carried out to determine the minimum concentrations of chlorhexidine that may restrain the conditioned candidal strains, in normoxia (normoxic MIC) and anoxia (anoxic MIC). The Mann-Whitney U test was used to evaluate the antimicrobial effect of chlorhexidine on C. albicans under normoxic and anoxic conditions (α = 0.05). Results: The normoxic MIC of chlorhexidine varied broadly from 150 to 1200 µg/mL, whereas its anoxic MIC varied narrower from 2.34 to 37.5 µg/mL. Regarding the origins of strains, no statistically significant differences (p > 0.05) were found. Conclusions: These results indicate that anoxic environmental conditions, compatible with periodontal pockets, tend to enhance C. albicans susceptibility to chlorhexidine.

Helicobacter pylori coinfection is a confounder, modulating mucosal inflammation in oral submucous fibrosis.

The oral cavity has been considered a potential reservoir for Helicobacter pylori (H pylori) , from where the organism causes recurrent gastric infections. Aim: With this case-control study we tried to evaluate the role of H pylori in the etiology of mucosal inflammation, a condition that compounds the morbid state associated with oral submucous fibrosis (OSF). Materials and Methods : Subjects ( n = 150) were selected following institutional regulations on sample collection and grouped into test cases and positive and negative controls based on the presence of mucosal fibrosis and inflammation. The negative controls had none of the clinical signs. All patients underwent an oral examination as well as tests to assess oral hygiene/periodontal disease status; a rapid urease test (RUT) of plaque samples was also done to estimate the H pylori bacterial load. We used univariate and mutivariate logistic regression for statistical analysis of the data and calculated the odds ratios to assess the risk posed by the different variables. Results : The RUT results differed significantly between the groups, reflecting the variations in the bacterial loads in each category. The test was positive in 52% in the positive controls (where nonspecific inflammation of oral mucosa was seen unassociated with fibrosis), in 46% of the test cases, and in 18% of the negative controls (healthy volunteers) (χ2 = 13.887; P < 0.01). A positive correlation was seen between the oral hygiene/periodontal disease indices and RUT reactivity in all the three groups. Conclusions: The contribution of the H pylori in dental plaque to mucosal inflammation and periodontal disease was significant. Logistic regression analysis showed gastrointestinal disease and poor oral hygiene as being the greatest risk factors for bacterial colonization, irrespective of the subject groups. A positive correlation exists between RUT reactivity and the frequency of mucosal inflammation.