A preliminary study on salivary microbiota of patients with laryngopharyngeal reflux / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the characteristics of salivary microbiota in patients with laryngopharyngeal reflux (LPR). Methods: A case-control study was applied to enroll 60 patients and healthy subjects who were outpatients of the Department of Otorhinolaryngology Head and Neck Surgery of the Eighth Medical Center of the PLA General Hospital from December 2020 to March 2021, including 35 males and 25 females, aged from 21 to 80 (33.75±11.10) years. Thirty patients with suspected laryngopharyngeal reflux were selected as study group and thirty healthy volunteers without pharyngeal symptoms were selected as control group. Their salivary samples were collected, and the salivary microbiota was detected and analyzed by 16S rDNA sequencing. SPSS 18.0 software was used for statistical analysis. Results: There was no significant difference in the diversity of salivary microbiota between the two groups. At the phylum classification level, the relative abundance of Bacteroidetes in the study group was higher than that in the control group[37.86(31.15, 41.54)% vs 30.24(25.51, 34.18)%,Z=-3.46,P<0.01]. And the relative abundance of Proteobacteria in the study group was lower than that in the control group [15.76(11.81, 20.17)% vs 20.63(13.98, 28.82)%, Z=-1.98,P<0.05]. At the genus level, the relative abundance of Prevotella, Lactobacillus, Parascardovia and Sphingobium in the study group was higher than that in the control group(Z values were-2.92, -2.69, -2.05, -2.31, respectively, P<0.05).And the relative abundance of Streptococcus, Cardiobacterium, Klebsiella and Uruburuella of study group was lower than that of control group(Z values were -2.43, -2.32, -2.17, -2.32, respectively, P<0.05). LEfSe difference analysis showed that there were 39 bacteria with significant differences between the two groups, including Bacteroidetes, Prevotellaceae and Prevotella, which were enriched in the study group, and Streptococcaceae, Streptococcus and other taxa, which were enriched in the control group(P<0.05). Conclusion: The changes of the microflora in the saliva between LPR patients and healthy people suggest that the dysbacteriosis might exist in LPR patients, which may play an important role in the pathogenesis and development of LPR.

Saliva microbiota and metabolite in individuals with caries or periodontitis / 中华口腔医学杂志

Objective: To detect and analyze the characteristics of oral microbiota in species composition, function and metabolism among caries, periodontitis and oral healthy individuals, hunting for the microbiome-derived biomarkers with specificity and sensitivity to estimate the occurrence of these two diseases. Methods: Saliva samples were collected from 10 patients with high caries risk [decayed-missing-filled teeth (DMFT)≥6, HC group] in Department of Endodontics, 10 patients with periodontitis of grade Ⅱ A-Ⅲ C (PG group) in Department of Periodontology and 10 oral healthy individuals (HH group) from School of Stomatology, The Fourth Military Medical University during from March 2022 to June 2022. A baseline examination was conducted on all participants, including their oral conditions of caries and periodontal health. Metagenomic sequencing (Illumina PE150 platform) and liquid chromatography-mass spectrometry were used to detect microorganisms and their metabolites in the samples respectively. The sequencing data were analyzed to obtain the information of microbial taxonomic composition, functional genes and metabolites in each group of samples. The basic oral conditions and saliva samples of subjects in each group were evaluated and collected by the same professional endodontist. Results: There were no significant difference in baseline characteristics such as age and sex among the subjects in each group (P>0.05). DMFT in HC group (9.0±1.7) was significantly higher than that in HH group (0) and PG group (0) (F=243.00, P<0.001). Sequencing data analysis showed that the taxonomic compositions of salivary microbiota in each group were mainly Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria and Fusobacteria at the phylum level, and Streptococcus, Neisseria, Rothia, Prevotella at the genus level. Differential analysis showed that, compared with the HH group, HC group and PG group had significant differences in taxonomic composition (P<0.05), and the most significant among them was Prevotella. At the species level, Prevotella pallens was the most significant change in HC group, and Porphyromonas gingivalis in PG group. Metabolite analysis showed that there were significant differences in metabolites between HC group and PG group. The results showed that, compared with the HH group, the most significant metabolite change was 3-hydroxy-1, 5-diphenylpentan-1-one in HC group (P=0.001) and N1 acetylspermine in PG group (P=0.002) respectively. Compared with the PG group, the metabolite of HC group with the most significant difference is D-glucosamine 6-phosphate (P=0.006). The metabolism gene function analysis showed that, the enrichment of carbohydrate metabolism related genes was highest in HC group, followed with HH group, and it was lowest in PG group. In addition, compared with the HH group, the abundance of functional genes related to glucose metabolism, such as ABC transporter and phosphotransferase system, were significantly decreased in PG group (P<0.05), but significantly increased in HC group (P<0.05). Conclusions: There is a significant correlation between the alternation of carbohydrate metabolism of salivary microbiota with the occurrence of caries and periodontitis. In the future, Prevotella pallens and 3-hydroxy-1, 5-diphenylpentan-1-one may be the potential biomarkers of caries; while Porphyromonas gingivalis and N1 acetylspermine work in the predictions of periodontitis.

Spectrophotometric Analysis of Streptococcus mutans Growth and Biofilm Formation in Saliva and Histatin-5 Relate to pH and Viscosity

ABSTRACT Objective: To analyze the ability of saliva in controlling the growth and the biofilm formation of Streptococcus mutans (S. mutans) as well as the effect of histatin-5 anti-biofilm relate to pH and saliva viscosity. Material and Methods: The S. mutans biofilm assayed by crystal violet 1% and its growth measured by spectrophotometer. The saliva viscosity was analyzed by viscometer, and pH of saliva was measured by pH meter. Results: Based on the optical density values, growth of S. mutans in saliva ranged <300 CFU/mL (0.1 nm) at concentrations of 25%, 12.5% and 6.25% for 24 hours. Whereas at the 48 h and 72 h period of incubation shown an increase in growth of S. mutans ranged 300-600 CFU/mL (0.2-0.36 nm). The inhibitory biofilm formation of S. mutans in saliva was significantly higher at concentrations of 12.5% and 6.25% at 24 h incubation times on a moderate scale, whereas the histatin-5 was effective to inhibit S. mutans biofilm on the 50 and 25 ppm. The saliva possessed a higher inhibitory of biofilm S. mutans than histatin-5 and good level viscosity (0.91-0.92 cP). Conclusion: The saliva was able to control the growth of S. mutans, and histatin-5 can inhibit the biofilm formation S. mutans. Furthermore, the saliva was also able to respond to the pH change with good viscosity of saliva.

Probable Association Between Oral Lichen Planus and presence of Helicobacter Pylori: a Preliminary Study in a Chilean Population / Probable Asociación entre Liquen plano Oral y la presencia de Helicobacter pylori: estudio Preliminar en Chile

ABSTRACT: Oral Lichen planus (OLP) is one of the main inflammatory diseases of the oral mucosa that is considered as a potentially malignant disorder. The exact pathogenesis of OLP remains to be completely understood. However, presence of bacteria has been associated to the inflammatory response observed in OLP. Particularly, Helicobacter pylori a major etiological agent of gastrointestinal inflammatory diseases and risk factor for gastric cancer, has been associated to Lichen planus. Here we studied a group of Chilean patients if there is any association between the presence of Helicobacter pylori and the clinical manifestation of OLP. We found a significant difference between the patients positive for H. pylori and the age of OLP diagnosis, suggesting that oral H. pylori might induce the disease at an earlier age. However, we could not confirm a statistically significance between the presence of the bacteria and OLP.

RESUMEN: Liquen Plano Oral (LPO) es una enfermedad inflamatoria de la mucosa oral considerada como desorden potencialmente maligno. La patogénesis exacta de LPO es desconocida. Sin embargo, se ha asociado la presencia de bacterias como responsables de la inflamación observada en LPO. Particularmente, Helicobacter pylori (H. pylori), agente etiológico principal de enfermedades inflamatorias gastrointestinales y factor de riesgo de cáncer gástrico, ha sido asociado con LPO. Se estudió la posible asociación entre H. pylori y manifestaciones clínicas de LPO en un grupo de pacientes Chilenos. Se encontró diferencia significativa entre los pacientes positivos para H. pylori y la edad de diagnóstico de LPO, sugiriendo que H. pylori podría inducir la enfermedad a temprana edad. Sin embargo, no se pudo confirmar significancia estadística entre la presencia de esta bacteria y la presencia de displasia en LPO.

Biomarcadores salivares no diagnóstico e no monitoramento de patologias orais e sistêmicas / Biomarcadores salivares en el diagnóstico y monitoreo de enfermedades bucales y sistémicas / Salivary biomarkers for the diagnosis and monitoring of oral and systemic diseases

RESUMO Introdução: A saliva humana é constituída de um vasto arsenal de produtos secretórios com imenso potencial informativo e útil para detecção de determinadas patologias. A facilidade de obtenção e a especificidade dos biomarcadores, faz desta uma importante ferramenta clínica como método diagnóstico. Objetivo: Verificar, através de uma revisão de literatura, a utilização da saliva como método diagnóstico para doenças orais e sistêmicas. Métodos: Foram selecionados artigos publicados em inglês, no banco de dados online PubMed com descritores "saliva", "biomarkers" e "diagnosis", entre o período de 2013 a setembro 2018. Foram incluídos artigos no idioma inglês, dos últimos 5 anos. Foram obtidos 245 artigos. O estudo se conteve em 17 destes. Análise e integração da informação: Foi notória a aplicabilidade da saliva como método diagnóstico para uma variedade de doenças, entre as quais se podem citar: doenças bacterianas como a cárie dentária, pela detecção do Streptococcus mutans e Lactobacillus spp.; doenças autoimunes como a Síndrome de Sjögren, indicada não apenas pela redução do fluxo salivar, como também pelo aumento na concentração de sódio, cloro, Imunoglobulina A (IgA), Imunoglobulina G (IgG), e Prostaglandina E2 (PGE2); doenças virais como no diagnóstico auxiliar do HIV 1 e 2 pela expressão de IgG e também no diagnóstico precoce de enfermidades malignas como o carcinoma de células escamosas e o câncer de mama, pela detecção de anticorpos frente à proteína p53 e pela presença de marcadores tumorais como o c-erbB-2. Conclusão: O diagnóstico molecular na cavidade oral por meio da saliva mostra ser um método simples, não invasivo e muito promissor para o diagnóstico e monitoramento de inúmeras patologias(AU)

RESUMEN Introducción: La saliva humana está constituida de un considerable arsenal de productos secretorios con inmenso potencial informativo y útil para la detección de determinadas enfermedades. La facilidad de obtención y la especificidad de los biomarcadores, hace de esta una importante herramienta clínica como método diagnóstico. Objetivo: Verificar, por medio de una revisión bibliográfica, la utilización de la saliva como método diagnóstico para enfermedades bucales y sistémicas. Métodos: Se seleccionaron artículos publicados en inglés, en la base de datos online PubMed con descriptores "saliva", "biomarkers" y "diagnosis", entre el período de 2013 a septiembre de 2018. Se incluyeron artículos en idioma inglés, de los últimos 5 años. Se obtuvieron 245 artículos. El estudio se circunscribió a 17. Análisis e integración de la información: Se observó la aplicabilidad de la saliva como método diagnóstico para una variedad de enfermedades, entre las que cabe citar: enfermedades bacterianas como la caries, por la detección del Streptococcus mutans y Lactobacillus spp.; las enfermedades autoinmunes como el síndrome de Sjögren, indicado no solo por la reducción del flujo salivar, sino también por el aumento en la concentración de sodio, cloro, Inmunoglobulina A (IgA), Inmunoglobulina G (IgG), y Prostaglandina E2 (PGE2); las enfermedades virales como el diagnóstico auxiliar del VIH 1 y 2 por la expresión de IgG y también el diagnóstico precoz de enfermedades malignas como el carcinoma de células escamosas y el cáncer de mama, por la detección de anticuerpos contra la proteína p53 y por la presencia de marcadores tumorales como el c-erbB-2. Conclusiones: El diagnóstico molecular en la cavidad bucal por medio de la saliva muestra ser un método simple, no invasivo y muy prometedor para el diagnóstico y monitoreo de innumerables enfermedades(AU)

ABSTRACT Introduction: Human saliva consists of a vast arsenal of secretory products with huge information potential useful for the detection of certain diseases. The easy availability and the specificity of biomarkers make them an important clinical tool as a diagnostic method. Objective: By means of a bibliographic review, verify the use of saliva as a diagnostic method for oral and systemic diseases. Methods: A bibliographic search was conducted in the online database PubMed for papers published in English from 2013 to September 2018, using the search terms "saliva", "biomarkers" and "diagnosis". Papers written in English in the last five years were selected. Of the 245 papers obtained, the study considered 17. Data analysis and integration: It was found that saliva may be used as a diagnostic method for a variety of diseases. These include bacterial diseases such as dental caries, by detection of Streptococcus mutans and Lactobacillus spp.; autoimmune diseases such as Sjögren's syndrome, indicated not only by the decrease in salivary flow, but also by the increase in the concentration of sodium, chlorine, immunoglobulin A (IgA), immunoglobulin G (IgG), and prostaglandin E2 (PGE2); viral diseases as in the auxiliary diagnosis of HIV 1 and 2 by IgG expression and also in the early diagnosis of malignant diseases such as squamous cell carcinoma and breast cancer by detection of antibodies against p53 protein and the presence of tumor markers such as c-erbB-2. Conclusions: Molecular diagnostic examination of the oral cavity using saliva has shown to be a simple, non-invasive and very promising method for the diagnosis and monitoring of numberless diseases(AU)

Protein Profiles of Individuals with Erosive Tooth Wear

Abstract Objective: To determine if protein profiles identified in saliva could be used to determine risk and severity of erosive tooth wear. Material and Methods: Three types of saliva sampling were performed to obtain saliva from 34 18-year old individuals that received regular dental check-ups, along with clinical status of the dentition and risk factor related to erosive tooth wear using the VEDE scale. Protein profiles in saliva were determined using electrophoresis and the calculation of the percentage of a specific band at a specific molecular weight in relationship to the total protein in that sample (% of total) using molecular weight standards. This quantification was repeated for each protein band across a range of molecular weights for each sample to test for association with erosive tooth wear status. Results: There were no differences in the number of detectable proteins sourced from the parotid gland, nor the unstimulated and stimulated whole saliva. Five out of the 34 individuals had no signs of erosive tooth wear despite an acidic diet and were more likely to have proteins with molecular weight smaller than 1 KDa (p=0.03). Conclusion: There is potential for the use of protein profiling to determine risks for erosive tooth wear.

Differences in Proteomic Profiles Between Caries Free and Caries Affected Children

Abstract Objective: To determine if there are differences in protein profiles in saliva depending if children of caries-free versus caries affected. Material and Methods: A cohort of 91 children with ages between 6 and 19 years, along clinical status of caries experience. Protein profiles in saliva were determined using electrophoresis and the calculation of the percentage of a specific band at a specific molecular weight in relationship to the total protein in that sample (% of total) using molecular weight standards. This quantification was repeated for each protein band across a range of molecular weights for each sample. Chi-square, Fisher's exact, and Student t-tests were used to compare the distributions between caries-free and caries affected children (α=0.05). Results: Histatin was more likely to be non-detectable or reduced in caries-free children (OR=7.56; 95% CI 1.62-35.13) and these children had on average one less gel band detected by the assay we used. Conclusion: We have found differences in proteins between caries affected and caries-free children, suggesting that this line of investigation holds the promise of providing new tools for caries management.

SARS-CoV-2 and Saliva as a Diagnostic Tool: A Real Possibility

Abstract Covid-19 is a respiratory disease caused by the SARS-CoV-2 virus. The high rate of contagion and the spread of the virus in the population make the early detection of the pathogen the means for the adequate targeting of infection control measures. WHO directs sample collection on upper respiratory specimens, including nasopharyngeal and oropharyngeal swab or wash in ambulatory patients, as well as lower respiratory specimens: sputum and/or endotracheal aspirate or bronchoalveolar lavage, in addition to citing blood and feces. Among the various sample collection methods, saliva has been investigated and reported as a potential source for diagnosis. Thus, we propose to evaluate the current scenario, based on recent publications on the perspective of detecting SARS-CoV-2 in saliva as a diagnostic method for Covid-19. The detection of SARS-CoV-2 through saliva seems to be very promising, although obstacles such as the technique and the location of the collection and the sample size of the research carried out so far may present a limitation for its use. The current scenario presents saliva as a reliable method for the detection of SARS-CoV-2, due to the ease of obtaining the samples, the possibility of self-collection, low cost because there is no need to use specific equipment, in addition to reducing the risk of transmission for health professionals.

Influence of salivary conditioning and sucrose concentration on biofilm-mediated enamel demineralization

Abstract The acquired pellicle formation is the first step in dental biofilm formation. It distinguishes dental biofilms from other biofilm types. Objective To explore the influence of salivary pellicle formation before biofilm formation on enamel demineralization. Methodology Saliva collection was approved by Indiana University IRB. Three donors provided wax-stimulated saliva as the microcosm bacterial inoculum source. Acquired pellicle was formed on bovine enamel samples. Two groups (0.5% and 1% sucrose-supplemented growth media) with three subgroups (surface conditioning using filtered/pasteurized saliva; filtered saliva; and deionized water (DIW)) were included (n=9/subgroup). Biofilm was then allowed to grow for 48 h using Brain Heart Infusion media supplemented with 5 g/l yeast extract, 1 mM CaCl2.2H2O, 5% vitamin K and hemin (v/v), and sucrose. Enamel samples were analyzed for Vickers surface microhardness change (VHNchange), and transverse microradiography measuring lesion depth (L) and mineral loss (∆Z). Data were analyzed using two-way ANOVA. Results The two-way interaction of sucrose concentration × surface conditioning was not significant for VHNchange (p=0.872), ∆Z (p=0.662) or L (p=0.436). Surface conditioning affected VHNchange (p=0.0079), while sucrose concentration impacted ∆Z (p<0.0001) and L (p<0.0001). Surface conditioning with filtered/pasteurized saliva resulted in the lowest VHNchange values for both sucrose concentrations. The differences between filtered/pasteurized subgroups and the two other surface conditionings were significant (filtered saliva p=0.006; DIW p=0.0075). Growing the biofilm in 1% sucrose resulted in lesions with higher ∆Z and L values when compared with 0.5% sucrose. The differences in ∆Z and L between sucrose concentration subgroups was significant, regardless of surface conditioning (both p<0.0001). Conclusion Within the study limitations, surface conditioning using human saliva does not influence biofilm-mediated enamel caries lesion formation as measured by transverse microradiography, while differences were observed using surface microhardness, indicating a complex interaction between pellicle proteins and biofilm-mediated demineralization of the enamel surface.

Efecto de una Pasta Dental Comercial Conteniendo Xilitol Sobre el Recuento de Streptococcus Mutans en Saliva de Gestantes: Ensayo Clínico Controlado Aleatorizado / Effect of a Commercial Toothpaste Containing Xylitol on the Count of Streptococcus Mutans in Saliva of Pregnant Women: Randomized Controlled Clinical Trial

RESUMEN: El objetivo del presente trabajo fue determinar el efecto de una pasta dental comercial conteniendo xilitol sobre el recuento de Streptococcus mutans en saliva de gestantes. El presente fue un ensayo clínico, a doble ciego, que se realizó en el Centro de Salud "José Olaya" (Chiclayo Perú), en enero de 2017. Se trabajó con una población muestral de 50 gestantes en el segundo trimestre que cumplieron con los criterios establecidos, distribuyéndolas en dos grupos: 25 gestantes usaron pasta dental con 10 % de xilitol y 25 gestantes usaron pasta dental sin xilitol. Se les tomó y procesó microbiológicamente una muestra de saliva antes del inicio del estudio y 14 días después del uso de las respectivas pastas. Se realizó el recuento de unidades formadoras de colonias (UFC) de Streptococcus mutans en saliva con una confiabilidad altamente significativa mediante el Coeficiente de Correlación Intraclase, calibración intra e interexaminador (1,000 y 0,999, respectivamente). El análisis de los datos se realizó mediante la prueba U de Mann-Whitney, considerando un nivel de significancia del 5 %. No se encontró diferencia entre las gestantes que emplearon pasta dental con xilitol en comparación con las que utilizaron pasta sin xilitol (p=0,062). Se concluyó que el efecto de la pasta dental comercial conteniendo xilitol es similar a una pasta sin xilitol sobre el recuento de Streptococcus mutans en saliva de gestantes.

ABSTRACT: The objective of the present study was to determine the effect of a commercial toothpaste containing xylitol on the counts of Streptococcus mutans in saliva of pregnant women. The present was a double-blind clinical trial performed at the "José Olaya" health Centre (Chiclayo Peru) in January 2017. We worked with a sample population of 50 pregnant in the second trimester that met the established criteria, distributing in two groups: 25 pregnant women used toothpaste with 10 % xylitol and 25 pregnant used toothpaste without xylitol. They were taken and microbiologically processed a sample of saliva before the start of the study and 14 days after the use of the respective toothpastes. The Colony-forming units (CFU) of Streptococcus mutans in saliva were counted with a highly significant reliability through the intraclass correlation coefficient, Intra-and Interexaminer calibration (1.000 and 0.999, respectively). Data analysis was performed using the Mann-Whitney U test, considering a 5 % significance level. No difference was found among the pregnant women who used xylitol toothpaste compared to those who used toothpaste without xylitol (p = 0,062). It was concluded that the effect of xylitol containing commercial toothpaste is similar to a toothpaste without xylitol on the count of Streptococcus mutans in the saliva of pregnant women.

Validation of an adherence assay to detect group mutans streptococci in saliva samples / Validación del Test de adherencia para el recuento de estreptococcos del grupo mutans en muestras de saliva

The aim of the present study was to validate and establish a cut off point and the predictive value of an adhesion test (AA-MSMG), as a microbiological method for evaluating cariogenic risk. The study is based on a variant (20% sucrose) of a selective medium descripted by Gold et al. (MSMG). This method differentiates mutans group streptococci (MGS) by exacerbating the production of insoluble extracellular polysaccharide which gives adhesion to surfaces such as glass, plastic and dental enamel. Caries assessment according to ICDAS was conducted in 154 patients (aged >21 years) who were attended at Preventive and Community Dentistry Department, School of Dentistry, University of Buenos Aires, Argentina, between August 2017 to August 2018. The study population was assigned to groups according to the presence/ absence of caries lesions: Group A: ICDAS lesion code = 0 (L=0) on all dental surfaces (n=23); and Group B: L>1 (n=131). After mouth-rinsing with distilled water, saliva samples were collected with fasting and hygiene protocol, and sent immediately to the Microbiological Diagnosis Laboratory, Microbiology Department, School of Dentistry, University of Buenos Aires. Samples were homogenized and serially diluted to the tenth. 100 pl of the dilutions were cultured in 25 cm² sterile plastic flasks containing 9.9 ml of modified selective medium described by Gold (MSMG-selective and differential medium). Cultures were incubated in an anaerobic atmosphere at 36 ± 1°C for 48 hours. The supernatants were eluted and the samples washed with sterile distilled water. Colony forming unit counts were performed by calibrated researchers (Kappa >0.75) using a stereoscopic microscope at 50X. Mutans group streptococci (MGS) counts ranged from 1x10(4) to 1x10(5) CFU/ml in group A, and were higher than 1x10(6) CFU/ml in Group B. Statically analysis of results (ROC) showed that the AAMSMG has a satisfactory predictive value (91%) and established a cutoff point in 1.68x10(5) UFC / ml. This would indicate that individuals whose MGS saliva counts are higher than the cutoff value would be 5 times more likely to develop dental caries. Adherence assay could be a useful microbiological predictor of caries risk.

El objetivo del presente estudio fue validar, establecer el punto de corte y valor predictivo de una técnica microbiológica para evaluar el nivel de estreptococos del grupo mutans en saliva. La técnica consiste en un test de adherencia que emplea un medio selectivo modificado (20% sacarosa) descripto por Gold et al. (TA-MSMG). Este método permite diferenciar a los estreptococos del grupo mutans (SGM) exacerbando la producción del polisacárido extracelular insoluble que le confiere adhesión a superficies como vidrio, plástico y esmalte dental. De acuerdo con los criterios de ICDAS se sembraron 154 salivas de pacientes mayores de edad, que asistieron al Servicio de Odontología Preventiva y Comunitaria de la Facultad de Odontología de la Universidad de Buenos Aires entre los meses de agosto de los años 2017 y 2018. La población estudiada fue asignada a dos grupos según la presencia / ausencia de lesiones de caries: Grupo A: código de lesión ICDAS = 0 (L = 0) en todas las superficies dentales (n = 23); y Grupo B: L> 1 (n = 131). Después de realizar un enjuague bucal con agua destilada, las muestras de saliva se recogieron según protocolo (ayuno de 4 horas y suspensión de higiene dental de 12 hs). Las muestras se remitieron de inmediato al Laboratorio de Diagnóstico Microbiológico, Departamento de Microbiología de la Facultad de Odontología, Universidad de Buenos Aires. Para su procesamiento, las muestras fueron homogeneizadas y diluidas al décimo. Se cultivaron 100 pl de las diluciones en botellas de plástico estériles de 25 cm² que contenían 9,9 ml de medio de Gold modificado (MSMG-20% sacarosa). Los cultivos se incubaron en atmósfera anaeróbica a 36 ± 1°C durante 48 horas. El sobrenadante se eluyó y las muestras se lavaron con agua destilada estéril. Los recuentos de unidades formadoras de colonias SGMfueron realizados por investigadores calibrados (Kappa >0.75) utilizando un microscopio estereoscópico a 50X. Los recuentos de SGM presentaron una variación entre 1x10(4)y 1x10(5) UFC/ml en el grupo A, mientras que en el Grupo B fueron superiores a 1x10(6) UFC/ml. El análisis estadístico de los resultados determinó una curva ROC que establece para el TA-MSMG un valor predictivo del 91% y un punto de corte en 1.68x10(5) UFC SGM / ml. Esto indicaría que los individuos cuyos recuentos en saliva de SGM sean superiores al valor de corte, tendrían 5 veces más posibilidades de desarrollar caries (5:1). Este método podría ser un instrumento útil al momento de evaluar (indicador microbiológico) el riesgo cariogénico del paciente.

Serão as bactérias cariogénicas o fator de risco maior para cáries dentárias em doentes com retocolite ulcerativa? / Are cariogenic bacteria the major risk factor to dental caries in patients with ulcerative colitis?

ABSTRACT BACKGROUND: High prevalence of dental caries in ulcerative colitis (UC) has been attributed to diet and changes in salivary environment. OBJECTIVE: We aimed to characterize the prevalence of dental caries, salivary flow rates, salivary buffering capacity and cariogenic bacteria counts of Mutans streptococci and Lactobacillus spp and to evaluate their relationship with drug therapy, disease activity and duration. METHODS: A cross-sectional study was performed with UC patients followed in a tertiary center. Participants were submitted to a questionnaire (including demographic data, oral hygiene, eating habits) and a clinical observation with assessment of plaque index and Decayed, Missing and Filled Teeth index. Unstimulated/stimulated saliva was collected. Medical records, disease activity (Partial Mayo Score) and disease duration were collected. Laboratory data included salivary flow rates, salivary buffering capacity (CRT® buffer) and cariogenic bacteria count (Mutans streptococci and Lactobacillus spp) in saliva using the CRT® bacteria test (results: high or low counts). RESULTS: Thirty UC patients were recruited. Oral hygiene routines were daily teeth brushing once or more (96.7%) and fluoride toothpaste (73.3%). Decayed, Missing and Filled Teeth index (mean 16.17±6.428) was not affected by the frequency of soft drinks, cakes, sweets and sugars between meals (P>0.2). Long-term disease showed a trend towards higher prevalence of caries (P=0.06). Most presented normal salivary flow rates, unstimulated (73.3%) and stimulated (60.0%), and high salivary buffering capacity (66.7%). Any association was found with age, gender, disease activity, disease duration and drug therapy. High Mutans streptococci and low Lactobacillus spp count were observed in 73.3% and 60% of patients, respectively. Patients with active disease (100%) and longer duration (88.9%) displayed higher Mutans streptococci count. CONCLUSION: The prevalence of dental caries observed in UC patients was significant and did not seem to be influenced by their eating habits. The high prevalence of Mutans streptococci count may be a major risk factor for dental caries and may be looked as part of the UC dysbiosis. Dental care of UC patients should be planned according with this microbiota variation.

RESUMO CONTEXTO: Uma elevada prevalência de cáries dentárias em doentes com retocolite ulcerativa (RCU) tem sido atribuída à dieta e alterações no ambiente salivar. OBJETIVO: Este estudo pretende caracterizar a prevalência de cáries dentárias, taxas de fluxo salivar, capacidade de tamponamento da saliva e contagem de bactérias cariogénicas, Streptococcus mutans e Lactobacillus spp e avaliar a sua relação com a terapêutica farmacológica e a atividade e duração da doença. MÉTODOS: Estudo transversal com doentes com RCU seguidos num centro terciário. Os participantes foram submetidos a um questionário (incluindo questões sobre dados sociodemográficos, higiene oral e hábitos alimentares), e a uma observação clínica com avaliação do índice de placa e índice Decayed, Missing and Filled Teeth. Amostras de saliva não estimulada e estimulada foram colhidas. Os registos clínicos, a atividade da doença (score parcial de Mayo) e a sua duração foram avaliados. Os dados laboratoriais incluíram taxas de fluxo salivar, capacidade de tamponamento da saliva (CRT® buffer) e a contagem de bactérias cariogénicas (Streptococcus mutans e Lactobacillus spp) utilizando o teste CRT® (resultado: contagem elevada ou reduzida). RESULTADOS: Foram recrutados 30 doentes com RCU. As rotinas de higiene oral incluíram a escovagem dos dentes uma ou mais vezes por dia (96,7%) e o uso de pasta com flúor (73,3%). O índice Decayed, Missing and Filled Teeth (média 16,17±6,428) não foi afetado pela frequência de ingestão de refrigerantes, bolos, doces e açucares entre as refeições (P>0,2). Doentes com doença de longa duração apresentaram uma tendência para maior prevalência de cáries dentárias (P=0,06). A maioria possuía taxas de fluxo salivar normais, quer de saliva estimulada (60%) quer de saliva não estimulada (73,3%), assim como taxas de tamponamento salivar elevadas (66,7%). Não foi encontrada associação entre estas características com a idade, género, atividade/duração da doença e tipo de terapêutica farmacológica. Contagens elevadas de Streptococcus mutans e reduzidas de Lactobacillus spp foram observadas em 73,3% e 60% dos doentes, respectivamente. Doentes com doença ativa e maior tempo de evolução apresentaram contagens superiores de Streptococcus mutans. CONCLUSÃO: A prevalência de cárie dentária observada em doentes com RCU foi significativa e não parece ser influenciada pelos hábitos alimentares. A prevalência aumentada de Streptococcus mutans foi o fator de risco maior para a cárie dentária e provavelmente constitui parte da conhecida disbiose oral associada à RCU. Os cuidados dentários a estes doentes deverão ter em consideração esta variação da microbiota.

Efectividad bactericida del diamino fluoruro de plata a diferente concentración sobre estreptococos cariogénicos en muestras de saliva y dentina de escolares: un estudio in vitro / Bactericidal effectveness of silver diamine fluoride in different concentrations on cariogenic Streptococcus in salivary and dentinal samples of schoolchildren: an in vitro study

La OMS y la FDI han publicado que entre el 60 y 90% de los escolares padecen caries. En nuestro país, el Sistema de Vigilancia Epidemiológica de Patologías Orales (SIVEPAB) 2012, reporta un 85% de caries a nivel nacional en población pediátrica. Los agentes anticariogénicos como el diamino y el fluoruro de plata son un tratamiento alentador, este agente puede actuar como bactericida o bacteriostático en función de su concentración y su capacidad para inhibir el crecimiento de estreptococos del grupo viridans, y por ende, de la caries. Problema: ¿Cuál es la efectividad bactericida del diamino fluoruro de plata (Saforide®) a diferente concentración sobre la microbiota cariogénica de escolares? Objetivo: Determinar la eficacia bactericida del diamino fluoruro de plata (DFP) a diferentes concentraciones en el crecimiento bacteriano de Streptococcus mitis, S. mutans y S. salivarius en muestras de saliva y dentina en escolares. Material y métodos: Se llevó a cabo un estudio experimental con una variable independiente, el efecto bactericida del diamino fluoruro de plata y se tomó el halo de inhibición como la dependiente. Se utilizaron medidas descriptivas como prueba de comparación y análisis de varianza usando post-hoc Tukey≠ con una confianza del 95%, y análisis de datos exploratorios. Resultados: Se analizaron 100 muestras, de las cuales 48.3% correspondió a S. mutans, 41.4% a S. salivarius y 10.3% a S. mitis, se obtuvo una mayor zona de inhibición para las tres bacterias al 38% mostrando una diferencia estadísticamente significativa 12% (p < 0.05). También se observó un efecto bacteriostático al 12%, no así para el 38%, donde se encontró un efecto bactericida Conclusión: Nuestros resultados sugieren que al 38% de la concentración hay un claro efecto bactericida en el grupo de estreptococos viridans y el 12% no se recomienda para la detención de caries debido al efecto bacteriostático (AU)

WHO and FDI have ruled that 60-90% of schoolchildren are affected by caries. In our country, the System of Epidemiological Surveillance of Oral Pathologies (SIVEPAB) (SIVEPAB) 2012. Report a rate of 85% of caries nationally in pediatric population. Anticariogenic diamino agents such as silver fluoride are an encouraging decrease in treatment for these high rates of tooth decay in our country, this agent can act as bactericidal or bacteriostatic based on their concentration and their ability to inhibit endogenous metalloproteinase (MMP-2, 8, 9). Problem: What will be the bactericidal effectiveness of silver diamine fluoride different concentration on cariogenic Streptococci saliva samples taken from school and dentin? Objective: Determine the bactericidal effectiveness Silver diamine fluoride (SDF) to different concentration on bacterial growth of Streptococcus mitis, S. mutans, and S. salivarius in saliva samples and dentin in school. Material and methods: An experimental study was conducted as an independent variable the bactericidal effect of silver diamine fluoride was taken as dependent inhibition halo. Descriptive measures were used as a comparison test and analysis of variance using Post-hoc Tukey with 95% confidence, and exploratory data analysis. Results: One hundred samples, of which 48.3% corresponded to S. mutans, 41.4% to S. salivarius and 10.3% to S. mitis, were analyzed, we obtained a larger zone of inhibition for all three organisms at 38% showing a statistically significant difference from 12% (p < 0.05). It was also observed that the 12% sample bacteriostatic effect, not to the concentration of 38% was found a bactericidal effect. Conclusion: Our results suggest that 38% concentration has a bactericidal effect on Streptococcus viridans group and 12% showed not recommended for the arrest or detention of dentine caries bacteriostatic effect (AU)

Early Detection of Bacillus anthracis From Saliva in Anticipation of a Bioterrorism Attack

Objective: To assess potential for early detection of oral infection by B. anthracis spores for preparedness of a bioterrorism attack. Material and Methods: The laboratory study used saliva with a range of initial anthrax concentrations, to compare detection by direct observation from conventional blood agar culture and by anthrax-specific PCR after a shorter culture in BHI broth. Three types of saliva were collected: stimulated saliva, unstimulated/whole saliva, and unstimulated/whole saliva with antibiotic treatment (for negative control). Using bivariate Kruskal-Wallis and Mann-Whitney tests for statistical analysis for factors that could affecting anthrax detection, significant differences between the test groups was assumed at p<0.05. Results: From unstimulated whole saliva heat shock treated at 62.50C, B. anthracis growth was detected with both methods. PCR detection from a BHI broth culture could shorten the time to diagnosis in comparison to conventional culture in blood agar. Conclusion: Saliva can provide useful samples for diagnosis of oropharyngeal anthrax. In comparison to conventional culture on blood agar, shorter-term culture in BHI broth provides potential for earlier detection and diagnosis.

Analysis of the Total Antioxidant Capacity of Saliva in Smokers

Objective: To analyze the total antioxidant capacity (TAoC) of saliva in smokers based on type of cigarette, duration and frequency of smoking. Material and Methods: 51 male smokers, aged 20-55 years were enrolled. Information regarding cigarette type and smoking duration and frequency was collected using a questionnaire. Unstimulated saliva samples were collected in the morning following fasting for 2 h, and TAoC was measured using a commercial kit. The data were evaluated through the independent t-test and Kruskal-Wallis test. Results: Mean TAoC for the consumption of Kretek cigarettes was 0.29 (±0.15) and for that of non-Kretek cigarettes was 0.36 (±0.10). Mean TAoC based on smoking duration was 0.31 (±0.14) for 5-10 years and 0.27 (±0.15) for >10 years. Median TAoC based on smoking frequency was 0.23 (0.11-0.44), 0.31 (0.06-0.64), and 0.27 (0.06-0.68) for 1-5, 6-10, and 11-20 cigarettes/day. Mean TAoC of the saliva from participants who consumed Kretek cigarettes was lower than that of the saliva from those who consumed non-Kretek cigarettes (p=0.3). Mean TAoC for a duration >10 years was lower than that for a duration of 5-10 years, although the difference between these two groups was not significant (p=0.4). Conclusion: There were tendencies of lower total antioxidant capacity on smokers with kretek type cigarettes, smoking duration >10 years and frequency of 1-5 cigarettes/day. This study indicates that the type, duration, and frequency of smoking may affect the salivary total antioxidant capacity.

Microbial species associated with dental caries found in saliva and in situ after use of self-ligating and conventional brackets

Abstract Objectives Enamel demineralization is among the main topics of interest in the orthodontic field. Self-ligating brackets have been regarded as advantageous in this aspect. The aim of this study was to evaluate the break homeostasis in the oral environment and the levels of microorganisms associated with dental caries among the different types of brackets. Material and Methods Twenty patients received two self-ligating brackets: In-Ovation®R, SmartClipTM, and one conventional GeminiTM. Saliva was collected before bonding (S0), 30 (S1) and 60 (S2) days after bonding. One sample of each bracket was removed at 30 and 60 days for the in situ analysis. Checkerboard DNA-DNA Hybridization was employed to evaluate the levels of microbial species as-sociated with dental caries. Data were evaluated by nonparametric Friedman and Wilcoxon tests at 5% significance level. Results The salivary levels of L. casei (p=0.033), S. sobrinus (p=0.011), and S. sanguinis (p=0.004) increased in S1. The in situ analyses showed alteration in S. mutans (p=0.047), whose highest levels were observed to the In-Ovation®R. Conclusions The orthodontic appliances break the salivary homeostasis of microorganisms involved in dental caries. The contamination pattern was different between self-ligating and conventional brackets. The In-Ovation®R presented worse performance considering the levels of cariogenic bacterial species.

Inflammatory markers in the saliva of cerebral palsy individuals with gingivitis after periodontal treatment

Abstract The aim of this study was to evaluate the effect of periodontal treatment on the salivary cytokine levels and clinical parameters of individuals with cerebral palsy (CP) with gingivitis. A non-randomized, clinical trial was conducted in individuals diagnosed with spastic CP. Thirty-eight individuals were enrolled in the study and were categorized according to gingival index scores between 0-1 or 2-3, assigned to groups G2 or G1, respectively. Periodontal treatment comprised oral hygiene instructions, conventional mechanical treatment and 0.12% chlorhexidine applied as an adjunct. Clinical parameters and saliva samples were collected at baseline and at the 15-day follow-up visit. Bleeding on probing and periodontal screening and recording were determined. Non-stimulated saliva samples were obtained, and the salivary flow rate, the osmolality and the levels of cytokines IL-1β, IL-6, IL-8, IL-10, TNF-α and IL-12p70 were evaluated by a cytometric bead array. The Wilcoxon test, the Mann-Whitney test, Spearman correlation analysis, Poisson regression analysis and an adjusted analysis were performed (α = 0.05). The groups differed significantly in periodontal clinical parameters at baseline and at follow-up. Salivary flow rate and osmolality were similar in both groups at both timepoints. However, TNF-α and IL-1β levels were higher in G1 than in G2 at baseline. Mechanical treatment resulted in improved clinical parameters for both groups. Furthermore, mechanical treatment resulted in a significant reduction in salivary IL-1β and IL-8 levels for both groups after treatment. Periodontal treatment performed in individuals with CP and gingivitis reduces the levels of TNF-α, IL-1β, IL-6 and IL-8.

Salivary Flow in Pediatric Cancer Patients Compared to Healthy Children and Adolescents

Abstract Objective: To verify differences between salivary flow in pediatric cancer patients before starting antineoplastic treatment and in healthy pediatric patients. Material and Methods: This is an observational, cross-sectional, paired study with sample of 120 children and adolescents (3-18 years). Thirty pediatric cancer patients were selected for convenience at "Napoleão Laureano" Hospital (G1). Another group was composed of 90 individuals attended at the School of Dentistry Clinics of the Federal University of Paraíba, matched by age (G2). Data collection was performed in two steps for both groups. Information regarding pediatric cancer patients was obtained by interview with parents / guardians and searching medical records, while in the other group by interview with parents / guardians. Saliva collection was performed using standard method in both groups: unstimulated salivary flow rate (USFR) being the mean volume expelled in 1 minute. Data were analyzed by the Shapiro-Wilk, Kolmogorov-Smirnov and Mann-Whitney tests (α = 5%) Results: Mean USFR for G1 and G2 was 0.52 mL / min and 0.66 mL / min, respectively (p>0.05) and, in both groups, significant difference was observed (p <0.05) between the mean USFR values of its subgroups, and values of adolescents being higher than those of children Conclusion: There is no difference in unstimulated salivary flow of pediatric cancer patients before starting antineoplastic treatment compared with healthy pediatric patients.

Streptococcus mutans detection in saliva and colostrum samples / Detecção de Streptococcus mutans em amostras de saliva e colostro

ABSTRACT Objective To detect Streptococcus mutans in colostrum and saliva of neonates and compare with its detection in saliva of mothers. Methods Forty-three healthy women, full-term gestations with no complications, submitted to elective Cesarean section, and their newborns were included in the study. Samples were investigated by polymerase chain reaction to detect S. mutans in genetic material from the samples. Results Approximately 16% of colostrum samples showed S. mutans , but not correlated with the presence of the bacteria in both samples of saliva. S. mutans was detected in 49 and 30% of saliva samples of mothers and neonates, respectively. There was a positive correlation in S. mutans detection between types of saliva. The number of maternal samples of saliva with detectable S. mutans was smaller in women receiving dental treatment during pregnancy. Tooth brushing, three times a day, influenced the detection of S. mutans in both the saliva and the colostrum. Conclusion Although maternal saliva may present S. mutans , few samples of colostrum present the bacteria. The presence of bacteria in neonate saliva may be related to contact before birth. Dental treatment and hygiene habits seem to influence the detection of S. mutans in samples of maternal saliva and colostrum.

RESUMO Objetivo Detectar Streptococcus mutans no colostro e na saliva de recém-nascido, e comparar com a detecção na saliva da mãe. Métodos Foram incluídos no estudo 43 mulheres saudáveis, com gestações a termo e sem complicações, que tiveram cesariana eletiva, e seus recém-nascidos. As amostras foram investigadas por reação de polimerase em cadeia para a detecção de S. mutans em material genético extraído das amostras. Resultados Cerca de 16% das amostras de colostro apresentaram S. mutans , não sem correlação com a presença das bactérias em ambas amostras de saliva. S. mutans foi detectado em 49 e 30% das amostras de saliva das mães e recém-nascidos, respectivamente. Houve correlação positiva na detecção de S. mutans entre os tipos de saliva. O número de amostras de saliva materna com S. mutans detectável foi menor nas mulheres que receberam tratamento odontológico durante a gravidez. A escovação três vezes ao dia influenciou na detecção do S. mutans tanto no saliva quanto no colostro. Conclusão Embora a saliva materna possa apresentar S. mutans , poucas amostras de colostro apresentam a bactéria. A presença de bactéria na saliva de neonatos pode estar relacionada ao contato antes do nascimento. O tratamento odontológico e os hábitos de higiene parecem influenciar na detecção de S. mutans em amostras de saliva e colostro maternos.

ERG11 gene polymorphisms and susceptibility to fluconazole in candida isolates from diabetic and kidney transplant patients

Abstract INTRODUCTION: Candidiasis is the most frequent opportunistic mycosis in humans and can cause mortality, particularly in immunodeficient patients. One major concern is the increasing number of infections caused by drug-resistant Candidas trains, as these cannot be efficiently treated with standard therapeutics. The most common mechanism of fluconazole resistance in Candida is mutation of ERG11, a gene involved in the biosynthesis of ergosterol, a compound essential for cell integrity and membrane function. METHODS: Based on this knowledge, we investigated polymorphisms in the ERG11 gene of 3 Candida species isolated from immunocompromised and immunocompetent patients. In addition, we correlated the genetic data with the fluconazole susceptibility profile of the Candida isolates. RESULTS: A total of 80 Candida albicans, 8 Candida tropicalis and 6 Candida glabrata isolates were obtained from the saliva of diabetic, kidney transplant and immunocompetent patients. Isolates were considered susceptible to fluconazole if the minimum inhibitory concentration was lower than 8 μg/mL. The amino acid mutations F105L, D116E, K119N, S137L, and K128T were observed in C. albicans isolates, and T224C and G263A were found in C. tropicalis isolates. CONCLUSIONS: Despite the high number of polymorphisms observed, the mutations occurred in regions that are not predicted to interfere with ergosterol synthesis, and therefore are not related to fluconazole resistance.