In Vitro Activity of Essential Oils Distilled from Colombian Plants against Candidaauris and Other Candida Species with Different Antifungal Susceptibility Profiles.

Multi-drug resistant species such as Candida auris are a global health threat. This scenario has highlighted the need to search for antifungal alternatives. Essential oils (EOs), or some of their major compounds, could be a source of new antifungal molecules. The aim of this study was to evaluate the in vitro activity of EOs and some terpenes against C. auris and other Candida spp. The eleven EOs evaluated were obtained by hydro-distillation from different Colombian plants and the terpenes were purchased. EO chemical compositions were obtained by gas chromatography/mass spectrometry (GC/MS). Antifungal activity was evaluated following the CLSI standard M27, 4th Edition. Cytotoxicity was tested on the HaCaT cell line and fungal growth kinetics were tested by time-kill assays. Candida spp. showed different susceptibility to antifungals and the activity of EOs and terpenes was strain-dependent. The Lippia origanoides (thymol + p-cymene) chemotype EO, thymol, carvacrol, and limonene were the most active, mainly against drug-resistant strains. The most active EOs and terpenes were also slightly cytotoxic on the HaCaT cells. The findings of this study suggest that some EOs and commercial terpenes can be a source for the development of new anti-Candida products and aid the identification of new antifungal targets or action mechanisms.

Molecular identification and genotyping of Streptococcus mutans from saliva samples of children in Medellin, Colombia / Identificación molecular y genotipificación de Streptococcus mutansde muestras de saliva de niños de Medellín, Colombia

Abstract Introduction and objective: Tooth decay is one of the most prevalent infectious diseases of the oral cavity. The aim of this study was to determine the prevalence of Streptoccocus mutans and its genotypes in saliva samples from 6 and 7 year old children with and without dental caries. Materials and methods: Forty seven saliva samples were obtained from 6-7 year-old children, randomly selected from a Public School in Medellin, Colombia. Oral examinations in order to determine the DMF-T Index (Decay, Missing and Filling Teeth) were carried out. Isolates were identified by the Streptoccocus sobrinus and Streptococcus mutans PCR, API 20STREP and 16S rRNA sequence analysis. Genotypes c, e, f and k were detected by PCR. Results: Prevalence of S. mutanswas 14.9%. Of the 47 saliva samples, 57.4% (27) corresponded to children with dental caries, and 8.5% (4) were positive for S. mutans genotype c, 2.1% (1 each) genotype f genotype k, and genotype cand k, respectively. Conclusion: Prevalence of S. mutanswas lower than in previous studies of Colombian children with similar demographic characteristics. Streptoccocusmutans'genotype c, fand kwere found in children with caries but not in the group without caries. This is the first report of S. mutans genotype kin Colombia; this genotype requires further study to clarify its relation with dental caries and cardiovascular disease in Colombia.

Resumen Introducción y objetivo: La caries dental es una de las enfermedades infecciosas de la cavidad oral más comunes en el mundo. El objetivo de este estudio fue determinar la prevalencia de Streptococcus mutansy sus genotipos en muestras de saliva de niños de 6 y 7 años, con y sin caries dental. Materiales y métodos: Cuarenta y siete (47) muestras de saliva fueron obtenidas de niños de 6-7 años, seleccionados aleatoriamente de una escuela pública en Medellín, Colombia. Se realizaron evaluaciones orales para determinar el índice COP-D (número de dientes permanentes cariados, obturados y perdidos). Los aislamientos fueron identificados mediante PCR para diferenciación de S. sobrinusy S. mutans, API-20STREP y análisis de ARNr 16S. Se determinaron mediante PCR los genotipos c e, fy ken las cepas identificadas como S. mutans. Resultados: La prevalencia de S. mutansfue 14,9%. De las 47 muestras de saliva, 57,4% (27) correspondieron a niños con caries dental, y 8,5 % (4), fueron positivas para S. mutans genotipo c, 2,1 % (1 cada una) genotipo f genotipo ky genotipo cy k, respectivamente. Conclusión: La prevalencia de S. mutansfue menor que la reportada en estudios previos de niños colombianos con características demográficas similares. Se encontró presencia de genotipos c, fy kde S. mutansen niños con caries dental, pero no en el grupo de niños sin caries dental. Este es el primer estudio en reportar la presencia del genotipo ken Colombia. Es necesario realizar más investigaciones en nuestro país para aclarar la relación de S. mutans genotipo k con caries dental y enfermedad cardiovascular.

[The physicochemical and microbiological characteristics of saliva during and after pregnancy]. / Características fisicoquímicas y microbiológicas de la saliva durante y después del embarazo.

OBJECTIVES: Identify the changes in some physiological and microbiological parameters in the saliva from a group of women during and after their pregnancies. METHODS: Stimulated whole saliva was collected from a cohort of 35 women during their pregnancy and afterwards to determine each sample's physicochemical (secretion rate, pH and buffer capacity) and microbiological characteristics (acidogenic bacteria count). RESULTS: The pH and buffer capacity of saliva during pregnancy were lower than after pregnancy. There were no statistically significant changes regarding S. mutans and Lactobacillus spp. count, but a tendency towards increased values during pregnancy was noted. CONCLUSIONS: Changes occurring in the saliva of pregnant women can lead to an increase of risk of suffering disease affecting one's oral health, such as caries, gingivitis and periodontal disease; this could be prevented by appropriate diagnosis and dental follow-up, including education regarding pregnant women's oral health.

Características fisicoquímicas y microbiológicas de la saliva durante y después del embarazo / The physicochemical and microbiological characteristics of saliva during and after pregnancy

Objetivo Identificar las variaciones producidas en algunos parámetros fisicoquímicos y microbiológicos de la saliva en un grupo de mujeres durante y después de la gestación. Materiales y Métodos Se recolectó saliva total estimulada en una cohorte de 35 mujeres embarazadas y después del parto. A cada muestra le fueron determinadas características fisicoquímicas (tasa de secreción, pH y capacidad amortiguadora) y microbiológicas (recuentos de bacterias acidogénicas). Resultados El pH y la capacidad amortiguadora de la saliva durante el embarazo fueron más bajos que en el posparto, mientras que la tasa de secreción salival fue más alta en el embarazo. Los recuentos de S. mutans y Lactobacillus spp. no presentaron cambios significativos estadísticamente, pero se observa una tendencia al aumento en los recuentos durante la gestación. Conclusión Los cambios ocurridos en la saliva de las gestantes pueden llevar a un aumento del riesgo de aparición de enfermedades bucales como caries, gingivitis y enfermedad periodontal, los cuales podrían disminuirse, mediante un adecuado diagnóstico y seguimiento odontológico, que incluya educación en salud oral a la gestante.

Objectives Identify the changes in some physiological and microbiological parameters in the saliva from a group of women during and after their pregnancies. Methods Stimulated whole saliva was collected from a cohort of 35 women during their pregnancy and afterwards to determine each sample's physicochemical (secretion rate, pH and buffer capacity) and microbiological characteristics (acidogenic bacteria count). Results The pH and buffer capacity of saliva during pregnancy were lower than after pregnancy. There were no statistically significant changes regarding S. mutans and Lactobacillus spp. count, but a tendency towards increased values during pregnancy was noted. Conclusions Changes occurring in the saliva of pregnant women can lead to an increase of risk of suffering disease affecting one's oral health, such as caries, gingivitis and periodontal disease; this could be prevented by appropriate diagnosis and dental follow-up, including education regarding pregnant women's oral health.

Comparison of the effect of two sugar-substituted chewing gums on different caries- and gingivitis-related variables: a double-blind, randomized, controlled clinical trial.

OBJECTIVE: The aim of this study was to compare the effect of two sugar-substituted chewing gums besides toothbrushing on different clinical, microbiological, and biochemical caries- and gingivitis-related variables. MATERIALS AND METHODS: The study was designed as a double-blind, randomized, controlled trial with three parallel arms. A total of 130 dental students, who volunteered after signing an informed consent, were randomly allocated to receive one of the following interventions: hexitol-sweetened gum containing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP), pentitol-sweetened gum containing no CPP-ACP, and control group with no gum. Subjects within the experimental groups chewed two gum pellets for 20 min three times a day after meals. The daily consumption level of both polyols was 6.0 g. Clinical examinations and salivary samplings were conducted at baseline and after 30 days of gum use. Pre- and post-intervention stimulated whole saliva samples were quantified for calcium/phosphate ionic concentration, total facultative bacterial load, Streptococcus mutans/Lactobacillus spp. counts, and Gram-negative percentage. RESULTS: A statistically significant reduction in visible plaque score was displayed in the hexitol/CPP-ACP gum group after the intervention when compared with baseline, but the order of the effect was in the same order as the differences between the groups at baseline. A similar tendency was seen in both the pentitol/non-CPP-ACP gum and control groups regarding total salivary facultative bacterial load and S. mutans count, but median values of these parameters were more significantly reduced in the pentitol/non-CPP-ACP gum group in comparison with those of the control group. Alterations of salivary Lactobacillus spp. were demonstrated only in the pentitol/non-CPP-ACP gum group. CONCLUSION: Although these findings might indicate that a 30-day protocol of daily chewing of pentitol-sweetened gum containing no CPP-ACP might have some a reducing effect on the salivary levels of facultative bacteria, S. mutans and Lactobacillus spp., there was only a marginal, if any, benefit from the chewing gums under study on some microbiological caries- and gingivitis-related variables. CLINICAL RELEVANCE: Taking into account that for transferring results into clinically relevant conclusions the findings need to be strong and consistent, adhering to single significant differences appears not appropriate. Hence, the clinical significance of chewing gums as an adjunctive tool for daily oral care remained questionable.

[Dental caries in young adults regarding saliva's microbiological and physical-chemical characteristics]. / Caries dental en adultos jóvenes en relación con características microbiológicas y fisicoquímicas de la saliva.

OBJECTIVE: Determining the relationship between saliva's physicochemical properties, cariogenic microorganism count, facultative anaerobic and gram-negative bacteria based on caries' experience in young adults. MATERIALS AND METHODS: Stimulated whole saliva was collected from 120 students aged 17 to 34 years old for analysing salivary flow rate, pH, calcium and phosphate ion concentration, lactic acid, cariogenic microorganism count and facultative and gram-negative bacteria. RESULTS: Salivary flow rate was included in the biological reference interval but was not found to be associated with caries; the same thing happened regarding lactic acid. A direct relationship was found between calcium and phosphate concentration and dental cavities. Streptococcus mutans was associated with white spot lesion whereas Lactobacillus spp., facultative anaerobic and gram-negative bacteria were associated with advanced cavities. CONCLUSIONS: Saliva's physicochemical and microbiological characteristics in the young adult group evaluated here were differentially related to caries in different degrees of progress.

Association of salivary levels of the bone remodelling regulators sRANKL and OPG with periodontal clinical status.

AIM: To determine the variations in salivary concentrations of sRANKL, osteoprotegerin (OPG) and its ratio, regarding the periodontal status. MATERIALS AND METHODS: Ninety-seven chronic periodontitis (CP) subjects and 43 healthy controls were selected. Periodontal status was assessed based on full-mouth clinical periodontal measurements. sRANKL and OPG salivary levels were analysed by ELISA. The association between these analytes and its ratio with CP was analysed individually and adjusted for confounding using a binary logistic regression model. RESULTS: sRANKL and sRANKL/OPG ratio were increased, whereas OPG was decreased in CP compared with healthy controls subjects. Although univariate analysis revealed a positive association of sRANKL salivary levels ≥6 pg/ml, OPG salivary levels ≤131 pg/ml and sRANKL/OPG ratio ≥0.062 with CP, after logistic regression analysis only the latter parameter was strongly and independently associated with disease status. Confounding and interaction effects of ageing and smoking habit on sRANKL and OPG levels could be noted. CONCLUSION: Although salivary concentrations of sRANKL, OPG and its ratio may act as indicators of the amount/extent of periodontal breakdown, the mutual confounding and synergistic biological interactive effects related to ageing and smoking habit of the susceptible host may also promote the tissue destruction in CP.

Salivary levels of matrix metalloproteinase (MMP)-9 and tissue inhibitor of matrix metalloproteinase (TIMP)-1: a pilot study about the relationship with periodontal status and MMP-9(-1562C/T) gene promoter polymorphism.

OBJECTIVE: Chronic periodontitis (CP) has been linked with an imbalance in the MMP-9/TIMP-1 ratio. A reasonable biologic explanation for this link is that the MMP-9 transcriptional activity can be modulated by MMP-9(-1562C/T) gene promoter polymorphism contributing to periodontal breakdown. This study aimed to assess the relationship between salivary MMP-9/TIMP-1 balance, MMP-9(-1562C/T) genotype and periodontal clinical status. DESIGN: Sixty-nine CP subjects and 54 healthy controls (HC) were selected. Periodontal status was assessed by criteria based on probing depth, clinical attachment level, extent, and severity of periodontal breakdown. Salivary levels of MMP-9 and TIMP-1 were analysed using ELISA and MMP-9(-1562C/T) genotype using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The association between salivary levels of MMP-9, TIMP-1 and MMP-9/TIMP-1 ratio with CP was assessed individually and adjusted for confounding using a binary logistic regression model. RESULTS: Significantly higher levels of both markers and their ratios were detected in the CP group in comparison to healthy controls. Synchronously, weak-to-moderate positive significant correlations between salivary biomarkers and clinical parameters were observed. After binary logistic regression analysis, salivary levels of MMP-9>20ngmL(-1), TIMP-1>64ngmL(-1) as well as MMP-9/TIMP-1 ratio >1 were independently associated with CP. Nevertheless, the MMP-9(-1562C/T) gene promoter polymorphism was not associated with the different degrees of chronic periodontitis and did not have influence on the salivary levels of biomarkers. CONCLUSION: The findings when considered within the limitations of this study may indicate that although a dominant expression of MMP-9 over TIMP-1 in saliva might reflect the periodontal clinical status, the functional polymorphisms in the promoter of the MMP-9(-1562C/T) gene from the Colombian population are not linked neither with significant salivary MMP-9 variations in these individuals nor periodontal clinical status.

Physicochemical salivary properties, Lactobacillus, Mutans Streptococci counts and early childhood caries in preschool children of Colombia

Dental caries is a multifactorial disease, in which saliva has an important role. Saliva properties affect the growth of cariogenic microorganisms, so variations below threshold levels are considered risk factors for the development of dental caries. Salivary tests and microbiological counts of cariogenic bacteria are often used for clinical purposes in order to reveal problems that could explain caries development and have been evaluated in multiple studies, but these studies have mainly been carried out on adults. Thus, the aim of this study was to identify physicochemical salivary properties, mutans streptococci and Lactobacillus counts among preschool children, and their relationship with ECC in a cross-sectional study in Medellin, Colombia. Caries was determined using ICDAS criteria for a sample of 201 preschool children. Stimulated saliva samples were processed for bacterial detection, mean flow rate, pH and buffer capacity calculation. Risk variables for ECC were assessed individually and adjusted using a binary logistic regression model. The results showed that 71.14% of study subjects had ECC. Flow rate, final pH and Buffer capacity increased with age, but none of them were related to the presence of ECC. Although OR analysis detected correlations between ECC, gender, age, dental eruption stage and Lactobacillus presence, binary logistic regression analysis only showed gender and Lactobacillus as strongly associated with ECC. It can be concluded that gender and Lactobacillus presence are key variables for the presence of ECC in these children; whilst age and dental eruption stage play a minor role. Further studies are required to clarify the role of mutans streptococci counts in this population and to establish normal salivary parameters as well as threshold levels for hyposalivation in preschool children, as results showed differences with normal parameters used for adults.

La caries dental es una enfermedad multifactorial en la cual la saliva tiene un impacto importante. Las propiedades de la saliva afectan el crecimiento de microorganismos cariogenicos y por lo tanto las variaciones por debajo de los niveles umbral son consideradas factor de riesgo para el desarrollo de caries. Las pruebas salivares y los recuentos de microorganismos cariogenicos son utilizadas con frecuencia en la clinica para identificar factores que puedan explicar el desarrollo de lesiones cariosas y han sido evaluados en multiples estudios, pero estos han sido desarrollados principalmente en adultos. El objetivo de este trabajo fue identificar propiedades fisicoquimicas de la saliva y recuentos de estreptococos del grupo mutans y Lactobacillus, en relacion con la Caries de la Infancia Temprana (CIT). Para esto, se realizo un estudio descriptivo transversal. Se registraron las lesiones de caries dental con el sistema ICDAS en 201 ninos pre-escolares de hogares infantiles de estrato socioeconomico medio-bajo. Se obtuvieron muestras de saliva estimulada, que fueron procesadas para detectar estreptococos del grupo mutans y Lactobacillus; se calculo la tasa de secrecion salivar, pH y capacidad Buffer. Se analizaron posibles variables indicadoras de riesgo para CIT, mediante el calculo de OR individuales por variable y OR ajustadas mediante un modelo de regresion logistica binario. Los resultados indicaron que el 71.14% de los ninos presentaban CIT. Las propiedades de la saliva mostraron un mejor comportamiento con la edad, con valores diferentes a los parametros definidos para poblaciones adultas, pero ninguna tuvo relacion con la enfermedad. El genero y la presencia de Lactobacillus mostraron asociacion fuerte en el analisis de regresion sugiriendolos como principales indicadores de riesgo para CIT en esta poblacion. Se requieren estudios prospectivos y analizar variaciones biologicas, para clarificar el papel de los estreptococos del grupo mutans y revisar los valores umbral para la evaluacion de la secrecion salivar en preescolares.

Screening for subgingival occurrence of gram-negative enteric rods in periodontally diseased and healthy subjects.

OBJECTIVE: This study aimed to determine whether the variations in the occurrence of gram-negative enteric rods as detected through a longitudinal follow up with three consecutive visits, may be associated with the periodontal clinical status. DESIGN: Clinical and demographic parameters from 63 untreated chronic periodontitis (CP) subjects and 45 healthy controls (HC) were analyzed. Subgingival plaque samples were obtained from 6 sites in each subject at baseline, 1-week, and 1-month visits and processed using culture and biochemical tests. Culture findings were categorized taking into account the detection frequency of gram-negative enteric rods as persistent presence, transient presence, or absence of enteric rods in any sampling time. RESULTS: Although transient presence of gram-negative enteric rods was more prevalent in CP subjects (16.7%) than HC subjects (9.3%), the difference was not significant (P>0.05). The majority of subjects showed a transient presence of gram-negative enteric rods at concentrations <2 x 10(2)CFU/mL. Persistent presence of gram-negative enteric rods was not observed in any subject through the entire study period. Moreover, differences in both the detection frequencies of individual species and the total number of gram-negative enteric rods were not statistically significant neither inter- nor intragroups. CONCLUSION: The findings of the present study could indicate that gram-negative enteric rods are merely transient microorganisms within the subgingival environment and suggest that the periodontal clinical status appeared not to be influenced by the presence of these species.

Physicochemical salivary properties, Lactobacillus, mutans streptococci counts and early childhood caries in preschool children of Colombia.

Dental caries is a multifactorial disease, in which saliva has an important role. Saliva properties affect the growth of cariogenic microorganisms, so variations below threshold levels are considered risk factors for the development of dental caries. Salivary tests and microbiological counts of cariogenic bacteria are often used for clinical purposes in order to reveal problems that could explain caries development and have been evaluated in multiple studies, but these studies have mainly been carried out on adults. Thus, the aim of this study was to identify physicochemical salivary properties, mutans streptococci and Lactobacillus counts among preschool children, and their relationship with ECC in a cross-sectional study in Medellín, Colombia. Caries was determined using ICDAS criteria for a sample of 201 preschool children. Stimulated saliva samples were processed for bacterial detection, mean flow rate, pH and buffer capacity calculation. Risk variables for ECC were assessed individually and adjusted using a binary logistic regression model. The results showed that 71.14% of study subjects had ECC. Flow rate, final pH and Buffer capacity increased with age, but none of them were related to the presence of ECC. Although OR analysis detected correlations between ECC, gender age, dental eruption stage and Lactobacillus presence, binary logistic regression analysis only showed gender and Lactobacillus as strongly associated with ECC. It can be concluded that gender and Lactobacillus presence are key variables for the presence of ECC in these children; whilst age and dental eruption stage play a minor role. Further studies are required to clarify the role of mutans streptococci counts in this population and to establish normal salivary parameters as well as threshold levels for hyprosalivation in preschool children, as results showed differences with normal parameters used for adults.