Anacardium excelsum phytochemical analysis and in vitro antimicrobial activity against oral cavity microorganisms.

Infections of the oral cavity have a broad microbial etiological profile that varies according to each microenvironment in the mouth. Such infections often require antimicrobial treatment, which can lead to the development of resistance. There is thus a need to find new therapeutic strategies based on natural plant-derived compounds. The aim of this study was to determine the phytochemical nuclei and the antimicrobial effect of Anacardium excelsum leaf and stem extracts, and of fractions derived from the leaf extract, against Streptococcus mutans ATCC 25175, Staphylococcus aureus ATCC 35548, Escherichia coli ATCC 25922, Enterococcus faecalis ATCC 19433 and Candida albicans ATCC 10231. The plant material was collected from the Quindío Botanical Garden (Calarcá, Quindío-Colombia), located at an altitude of 1500 meters above sea level. Hydroalcoholic extracts of A. excelsum leaves and stems, and fractions of the hydroalcoholic leaf extract, were obtained by percolation extraction. Phytochemical nuclei were identified by thin layer chromatography. The antimicrobial activity of the extracts and fractions (at concentrations of 2, 5, 10, 20 and 40 mg / ml) against the five ATCC reference strains was evaluated using the well diffusion technique on Mueller-Hinton agar. The leaf extract showed no antimicrobial activity against E. coli, but it did show antimicrobial activity against S. mutans, S. aureus, E. faecalis and C. albicans, at a concentration of 10 mg/ml, with zones of inhibition of 9 to 11 mm. The ethyl acetate and acetone fractions obtained from A. excelsum leaf extract had greatest antimicrobial activity at 10 mg/ml. In conclusion, (1) the A. excelsum leaf extract, and the ethyl acetate and acetone fractions obtained from the leaf extract, had the greatest antimicrobial activity on all the study microorganisms, and (2) the phytochemical nuclei in the fractions (ethyl acetate and acetone) were found to contain phenolic-type compounds, tannins, triterpene-type terpenes and steroidal-type terpenes, which might explain the antimicrobial activity observed.

Las infecciones de la cavidad bucal se caracterizan por presentar un amplio perfil etiológico microbiano que varía de acuerdo a cada microambiente en boca. En muchos casos estas infecciones requieren tratamiento antimicrobiano que puede conducir al desarrollo de resistencia. Estos hechos en conjunto generan la necesidad de buscar nuevas estrategias terapéuticas, provenientes de compuestos naturales derivados de plantas. El objetivo de este estudio fue determinar los núcleos fitoquímicos y el efecto antimicrobiano de extractos de hojas y tallos, y de fracciones derivadas del extracto de hojas de Anacardium excelsum contra Streptococcus mutans ATCC 25175, Staphylococcus aureus ATCC 35548, Escherichia coli ATCC 25922, Enterococcus faecalis ATCC 19433 y Candida albicans ATCC 10231. El material vegetal se colectó del Jardín Botánico del Quindío (Calarcá, Quindío-Colombia), situado a una altura de 1500 msnm. Se obtuvieron extractos hidroalcohólicos de hojas y tallos, y fracciones a partir del extracto hidroalcohólico de hojas de A. excelsum mediante extracción por percolación. La identificación de los núcleos fitoquímicos fue realizado por cromatografía en capa delgada. La evaluación de la actividad antimicrobiana de extractos y fracciones, en concentraciones de 2, 5, 10, 20 y 40 mg/ml, frente a las 5 cepas de referencia ATCC, se realizó mediante la técnica de difusión con aplicación en pozo sobre agar Mueller-Hinton. El extracto de hojas no presentó actividad antimicrobiana sobre E. coli pero si sobre S. mutans, S. aureus, E. faecalis y C. albicans, en una concentración de 10 mg/ml, con halos de inhibición desde 9 a 11 mm, y las fracciones de acetato de etilo y acetona obtenidas del extracto de hojas de A. excelsum, presentaron mayor actividad antimicrobiana en una concentración de 10 mg/ml. En conclusión, 1. El extracto de hojas y las fracciones de acetato de etilo y acetona obtenidas del extracto de hojas de A. excelsum presentaron mayor actividad antimicrobiana sobre todos los microorganismos en estudio; 2. La evaluación de los núcleos fitoquímicos en las fracciones acetato de etilo y de acetona mostraron compuestos de tipo fenólico, taninos, terpenos del tipo triterpenos y terpenos del tipo esteroidal, que posiblemente expliquen la actividad antimicrobiana obtenida.

Anacardium excelsum phytochemical analysis and in vitro antimicrobial activity against oral cavity microorganisms / Análisis fitoquímico y actividad antimicrobiana in vitro de Anacardium excelsum sobre microorganismos de cavidad bucal

ABSTRACT Infections of the oral cavity have a broad microbial etiological profile that varies according to each microenvironment in the mouth. Such infections often require antimicrobial treatment, which can lead to the development of resistance. There is thus a need to find new therapeutic strategies based on natural plant-derived compounds. The aim of this study was to determine the phytochemical nuclei and the antimicrobial effect of Anacardium excelsum leaf and stem extracts, and of fractions derived from the leaf extract, against Streptococcus mutans ATCC 25175, Staphylococcus aureus ATCC 35548, Escherichia coli ATCC 25922, Enterococcus faecalis ATCC 19433 and Candida albicans ATCC 10231. The plant material was collected from the Quindío Botanical Garden (Calarcá, Quindío-Colombia), located at an altitude of 1500 meters above sea level. Hydroalcoholic extracts of A. excelsum leaves and stems, and fractions of the hydroalcoholic leaf extract, were obtained by percolation extraction. Phytochemical nuclei were identified by thin layer chromatography. The antimicrobial activity of the extracts and fractions (at concentrations of 2, 5, 10, 20 and 40 mg / ml) against the five ATCC reference strains was evaluated using the well diffusion technique on Mueller-Hinton agar. The leaf extract showed no antimicrobial activity against E. coli, but it did show antimicrobial activity against S. mutans, S. aureus, E. faecalis and C. albicans, at a concentration of 10 mg/ml, with zones of inhibition of 9 to 11 mm. The ethyl acetate and acetone fractions obtained from A. excelsum leaf extract had greatest antimicrobial activity at 10 mg/ml. In conclusion, (1) the A. excelsum leaf extract, and the ethyl acetate and acetone fractions obtained from the leaf extract, had the greatest antimicrobial activity on all the study microorganisms, and (2) the phytochemical nuclei in the fractions (ethyl acetate and acetone) were found to contain phenolic-type compounds, tannins, triterpene-type terpenes and steroidal-type terpenes, which might explain the antimicrobial activity observed.

RESUMEN Las infecciones de la cavidad bucal se caracterizan por presentar un amplio perfil etiológico microbiano que varía de acuerdo a cada microambiente en boca. En muchos casos estas infecciones requieren tratamiento antimicrobiano que puede conducir al desarrollo de resistencia. Estos hechos en conjunto generan la necesidad de buscar nuevas estrategias terapéuticas, provenientes de compuestos naturales derivados de plantas. El objetivo de este estudio fue determinar los núcleos fitoquímicos y el efecto antimicrobiano de extractos de hojas y tallos, y de fracciones derivadas del extracto de hojas de Anacardium excelsum contra Streptococcus mutans ATCC 25175, Staphylococcus aureus ATCC 35548, Escherichia coli ATCC 25922, Enterococcus faecalis ATCC 19433 y Candida albicans ATCC 10231. El material vegetal se colectó del Jardín Botánico del Quindío (Calarcá, Quindío-Colombia), situado a una altura de 1500 msnm. Se obtuvieron extractos hidroalcohólicos de hojas y tallos, y fracciones a partir del extracto hidroalcohólico de hojas de A. excelsum mediante extracción por percolación. La identificación de los núcleos fitoquímicos fue realizado por cromatografía en capa delgada. La evaluación de la actividad antimicrobiana de extractos y fracciones, en concentraciones de 2, 5, 10, 20 y 40 mg/ml, frente a las 5 cepas de referencia ATCC, se realizó mediante la técnica de difusión con aplicación en pozo sobre agar Mueller-Hinton. El extracto de hojas no presentó actividad antimicrobiana sobre E. coli pero si sobre S. mutans, S. aureus, E. faecalis y C. albicans, en una concentración de 10 mg/ml, con halos de inhibición desde 9 a 11 mm, y las fracciones de acetato de etilo y acetona obtenidas del extracto de hojas de A. excelsum, presentaron mayor actividad antimicrobiana en una concentración de 10 mg/ml. En conclusión, 1. El extracto de hojas y las fracciones de acetato de etilo y acetona obtenidas del extracto de hojas de A. excelsum presentaron mayor actividad antimicrobiana sobre todos los microorganismos en estudio; 2. La evaluación de los núcleos fitoquímicos en las fracciones acetato de etilo y de acetona mostraron compuestos de tipo fenólico, taninos, terpenos del tipo triterpenos y terpenos del tipo esteroidal, que posiblemente expliquen la actividad antimicrobiana obtenida.

Antimicrobial Activity of Piper marginatum Jacq and Ilex guayusa Loes on Microorganisms Associated with Periodontal Disease.

BACKGROUND: Chronic periodontitis is a multifactorial infectious disease, where multiple bacteria, such as Porphyromonas gingivalis, Prevotella intermedia, and Fusobacterium nucleatum are implicated. The main purpose of researching natural products is to find substances or compounds with antimicrobial activity. AIM: The objective of this work was to determine antimicrobial activity from extracts and obtained fractions from Piper marginatum Jacq and Ilex guayusa Loes on P. gingivalis ATCC 33277, F. nucleatum ATCC 25586, and P. intermedia ATCC 25611. METHODS: Total ethanol extracts were obtained from both plants. Fractions were obtained from total ethanol extracts with amberlite as a stationary phase employing hexane, acetone, and ethanol-water as solvents. Qualitative and quantitative phytochemical characterization was performed on total ethanol extracts from both plants. Antimicrobial activity from total ethanol extracts and fractions from both plants were evaluated on P. gingivalis ATCC 33277, F. nucleatum ATCC 25586, and P. intermedia ATCC by the well diffusion method with Wilkins-Chalgren agar. RESULTS: Piper marginatum Jacq total ethanol extract presented antimicrobial activity against all three bacteria, whereas Ilex guayusa Loes was only efficient against P. gingivalis ATCC 33277 and P. intermedia ATCC 25611, with inhibition halos from 9.3 to 30 mm. Ilex guayusa Loes obtained fractions presented antimicrobial activity against all three microorganisms evaluated, with inhibition halos ranging from 9.7 to 18.7 mm. In regards to Piper marginatum Jacq fractions, inhibition halos were between 8.3 and 19 mm, against all three microorganisms evaluated; only hexane fraction did not present antimicrobial activity against F. nucleatum ATCC 25586. CONCLUSION: Piper marginatum Jacq and Ilex guayusa Loes total ethanol extracts and fractions presented outstanding antimicrobial activity against P. gingivalis ATCC 33277, P. intermedia ATCC 25611, and F. nucleatum ATCC 25586.

Presence and count of S. mutans in children with dental caries: before, during and after a process of oral health education.

Dental caries is an infectious, multifactorial, localized, transmissible process that leads to the destruction of hard dental tissue. Streptococcus mutans is considered to be the main microorganism associated with its development. The aim of this study was to determine presence and count of S. mutans in saliva samples from children with dental caries before and after an educational process including interviews, lessons, lectures, educational workshops and recreational activities on the importance of oral care and hygiene. Twenty-three 3- to 6-year-old schoolchildren provided 3 unstimulated saliva samples: one before the educational process, one at 3 months and one at 6 months into the educational process. The samples were serially diluted and plated on Mitis Salivarius agar supplemented with bacitracin and 20% sucrose, and incubated anaerobically for 2 days at 37º C. Presumptive S. mutans isolates were identified with biochemical tests. Before the beginning of the educational process, and at 3 and 6 months into the educational process, S. mutans was found, respectively, in 22 (95.6%), 15 (65.2%) and 10 (43.5%) of the 23 children. The S. mutans count was reduced by 64.8% and 86.6% at 3 and 6 months, respectively, compared to the levels found before the educational process. These results indicate that educational intervention produced a significant reduction in S. mutans levels in the saliva of children with dental caries at 3 and 6 months into the educational process.

La caries dental es un proceso infeccioso multifactorial, localizado y transmisible que se caracteriza por la destrucción del tejido dental duro. Streptococcus mutans es considerado el principal microorganismo asociado al desarrollo de esta enfermedad. El objetivo de este estudio fue determinar la presencia y recuento de S. mutans en saliva de niños con caries dental antes y después de un proceso educativo. Con este fin se tomó saliva no estimulada de 23 niños con caries dental pertenecientes a un centro educativo con edades de 3 a 6 años. En todos los niños se tomaron 3 muestras de saliva: antes del proceso educativo y a los 3 y 6 meses de iniciado el proceso educativo. El proceso educativo consistió en entrevistas, enseñanzas, conferencias, talleres educativos y actividades lúdicas sobre la importancia del cuidado e higiene oral. Después de su recolección, las muestras de saliva fueron serialmente diluidas y sembradas en Agar Mitis Salivarius con bacitracina y sacarosa al 20%. Los medios de cultivo sembrados se incubaron en anaerobiosis durante 2 días a 37°C y los aislamientos presuntivos de S. mutans se identificaron con pruebas bioquímicas. Antes del inicio del proceso educativo, a los 3 y 6 meses de iniciado el proceso educativo se encontró S. mutans, respectivamente, en 22 de los 23 niños (95.6%), en 15 de los 23 niños (65.2%) y en 10 de los 23 niños (43.5%). En cuanto al recuento de S. mutans, se encontró una reducción de 64.8 % y 86.6% a los 3 y 6 meses, respectivamente, en comparación a los niveles encontrados antes del inicio del proceso educativo. En conclusión, los resultados indican que la intervención educativa realizada produjo una reducción significativa en los niveles de S. mutans en saliva de niños con caries dental después de 3 y 6 meses de iniciado el proceso educativo.

Genotipificación, niveles de expresión y estado físico del virus del papiloma humano en pacientes colombianos con cáncer de células escamosas en la cavidad oral / Genotyping, levels of expression and physical status of human papilloma virus in oropharyngeal squamous cell carcinoma among Colombian patients

Introducción. Uno de los factores de riesgo del carcinoma de células escamosas en la cavidad oral es la infección por el virus del papiloma humano (HPV), cuyas prevalencias dependen de la región geográfica. Objetivo. Identificar los tipos del virus del papiloma humano más frecuentes en el cáncer de la cavidad bucal, sus niveles de expresión y el estado físico del genoma viral. Materiales y métodos. Se seleccionaron 46 pacientes que asistían a los servicios de cirugía de cabeza y cuello en Bogotá, Manizales y Bucaramanga. El examen histopatológico de las muestras incluidas en el estudio demostró la presencia de carcinoma de células escamosas en la cavidad oral en todas ellas. Se extrajo el ADN para genotipificar el virus y determinar el estado físico de su genoma, y el ARN para determinar los transcritos virales mediante reacción en cadena de la polimerasa en tiempo real. Resultados. La prevalencia del virus del papiloma humano en los tumores fue de 21,74% (n=10) y el tipo viral más frecuente fue el HPV-16 (nueve casos). La expresión viral del HPV-16 fue baja (una de 11 copias) y el estado físico predominante fue el mixto (ocho casos), con prevalencia de la disrupción en el sitio de unión de E1 y E2 (2525 a 3720 nucleótidos). Conclusión. En los pacientes con carcinoma de cavidad oral incluidos en este trabajo, la frecuencia del virus del papiloma humano fue relativamente baja (21,7 %) y el tipo viral más frecuente fue el HPV-16, el cual se encontró en forma mixta y con baja expresión de E7 , lo cual puede ser indicativo de un mal pronóstico para el paciente.

Introduction: One of the risk factors for squamous cell oropharyngeal carcinoma is infection with the human papilloma virus (HPV), with prevalences that vary depending on the geographical region. Objective: To identify the most frequent HPV viral types in oropharyngeal cancer, the levels of expression and the physical condition of the viral genome. Materials and methods: Forty-six patients were included in the study from among those attending head and neck surgical services in the cities of Bogotá, Manizales and Bucaramanga. In the histopathological report all study samples were characterized as oropharyngeal squamous cell carcinoma. DNA extraction was subsequently performed for HPV genotyping and to determine the physical state of the viral genome, as well as RNA to determine viral transcripts using real-time PCR. Results: HPV prevalence in tumors was 21.74% (n=10) and the most common viral type was HPV-16 (nine cases). Viral expression for HPV-16 was low (one of 11 copies) and the predominant physical state of the virus was mixed (eight cases), with disruption observed at the E1 - E2 binding site (2525 - 3720 nucleotides). Conclusion: The prevalence of HPV associated with oropharyngeal carcinoma among the Colombian study population was 21.7%, which is relatively low. The most frequent viral type was HPV-16, found in a mixed form and with low expression of E7 , possibly indicating a poor prognosis for these patients.

Biotipos y susceptibilidad antimicrobiana de S. mutans en niños con y sin caries dental / Biotypes and antimicrobial susceptibility of S. mutans in children with and without dental caries

Objetivos: investigar los biotipos de S. mutans aislados de niños con y sin caries dental y determinar su susceptibilidad antimicrobiana. Métodos: este estudio observacional descriptivo incluyó 206 niños entre los 3 y 5 años de dos preescolares en Bogotá, D. C., Colombia. Después de su aislamiento en agar Mitis Salivarius Bacitracina, las cepas S. mutans se identificaron por pruebas bioquímicas y se biotipificaron por el sistema api-ZYM. En todos los aislamientos se determinó por el método de dilución en agar la concentración inhibitoria mínima (CIM) de Penicilina, Amoxicilina, Cefazolina, Eritromicina, Clindamicina, Imipenem, Vancomicina y Teicoplanina. Resultados: Se encontró S. mutans en 30 de los 79 niños sin caries dental (38%) y en 56 de los 127 niños con caries (44,1%). Los niños con caries presentaron un recuento superior de S. mutans en comparación con los niños sin caries (p < 0,05). En total se identificaron 121 cepas de S. mutans en los 86 niños: 43 cepas en los 30 niños sin caries dental y 78 en los 56 niños con caries dental. Las 121 cepas fueron agrupadas en 38 biotipos: 24 en el grupo de caries y 14 en el grupo sin caries. En el grupo de caries los biotipos más frecuentes fueron el XV, XI, XII y XVII, respectivamente, con 26, 12, 11 y 4 cepas y en el grupo sin caries los biotipos más frecuentes fueron el XXVI, XX y XXXVI, respectivamente, con 14, 8 y 7 cepas. Todos los biotipos fueron altamente sensibles a los antimicrobianos evaluados; el 50 y 90% de las cepas S. mutans fueron inhibidas, respectivamente, por concentraciones menores a 0,12 y 1 µg/ml para todos los antimicrobianos estudiados. Para la penicilina, eritromicina e imipenem se obtuvieron los valores promedios más bajos de CIMs. Conclusiones: en la población objeto de estudio se encontró una gran diversidad biotípica, los aislamientos fueron altamente susceptibles a los antimicrobianos evaluados y en cada grupo de pacientes se identificaron patrones únicos que podrían indicar una colonización específica.

Objectives: The main objective was to investigate the biotypes of S. mutans isolated from children with and without dental caries and determine its antimicrobial susceptibility. Methods: this descriptive observational study included 206 children aged 3 to 5 years of two preschool institutions in Bogota-Colombia. After isolation on agar Mitis Salivarius Bacitracin, S. mutans strains were identified by biochemical tests and biotyped by the api-ZYM system. The minimal inhibitory concentrations (MICs) of the S. mutans isolates were evaluated against penicillin, amoxicillin, cefazolin, erythromycin, clindamycin, imipenem, vancomycin and teicoplanin by an agar dilution method. Results: S. mutans was found in 30 of the 79 children without dental caries (38%) and in 56 of the 127 children with caries (44.1%). Children with cavities had a higher count of S. mutans compared to children without caries (p < 0.05). A total of 121 strains of S. mutans were identified in 86 children: 43 strains in 30 children with dental caries and 78 in the 56 children with dental caries. The 121 strains were grouped into 38 biotypes: 24 in the group caries and 14 in the group without caries. In the group caries the most common biotypes were the XV, XI, XII and XVII, respectively, with 26, 12, 11 and 4 strains and the group caries the most common biotypes were the XXVI, XX and XXXVI, respectively, with 14, 8 and 7 strains. All biotypes were highly sensitive to all antimicrobials tested; 50 and 90% of S. mutans strains were inhibited by using concentrations lower than 0.12 and 1 µg/ml, respectively, for all antibiotics studied. The lowest MICs average values were obtained for penicillin, erythromycin and imipenem. Conclusions: In the study population a wide variety of biotypes was found, isolates were highly sensitive to antimicrobials evaluated and in each clinical group were identified unique patters that could indicate a specific colonization.

Genotyping, levels of expression and physical status of human papilloma virus in oropharyngeal squamous cell carcinoma among Colombian patients.

INTRODUCTION: One of the risk factors for squamous cell oropharyngeal carcinoma is infection with the human papilloma virus (HPV), with prevalences that vary depending on the geographical region.  OBJECTIVE: To identify the most frequent HPV viral types in oropharyngeal cancer, the levels of expression and the physical condition of the viral genome.  MATERIALS AND METHODS: Forty-six patients were included in the study from among those attending head and neck surgical services in the cities of Bogotá, Manizales and Bucaramanga. In the histopathological report all study samples were characterized as oropharyngeal squamous cell carcinoma. DNA extraction was subsequently performed for HPV genotyping and to determine the physical state of the viral genome, as well as RNA to determine viral transcripts using real-time PCR.  RESULTS: HPV prevalence in tumors was 21.74% (n=10) and the most common viral type was HPV-16 (nine cases). Viral expression for HPV-16 was low (one of 11 copies) and the predominant physical state of the virus was mixed (eight cases), with disruption observed at the E1 - E2 binding site (2525 - 3720 nucleotides).  CONCLUSION: The prevalence of HPV associated with oropharyngeal carcinoma among the Colombian study population was 21.7%, which is relatively low. The most frequent viral type was HPV-16, found in a mixed form and with low expression of E7, possibly indicating a poor prognosis for these patients.

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

Chronic periodontitis is a multifactorial infectious disease associated with Gram-negative strict anaerobes which are immersed in the subgingival biofilm. Porphyromonas gingivalis, an important periodontal pathogen, is frequently detected in patients with chronic periodontitis. Although isolates of P. gingivalis tend to be susceptible to most antimicrobial agents, relatively little information is available on its in vitro antimicrobial susceptibility. The aim of this study was to determine the frequency of P. gingivalis in patients with chronic periodontitis and to assess antimicrobial susceptibility in terms of minimum inhibitory concentration (MIC) of clinical isolates to metronidazole and tetracycline. A descriptive, observational study was performed including 87 patients with chronic periodontitis. Samples were taken from the periodontal pocket using paper points, which were placed in thioglycollate broth. Samples were incubated for 4 hours at 37°C in anaerobic conditions and finally replated on Wilkins-Chalgren anaerobic agar (Oxoid). Bacteria were identified using the RapIDTMANAII system (Remel) and antimicrobial susceptibility was determined with the M.I.C. Evaluator test (MICE, Oxoid). P. gingivalis was identified in 30 of the 87 patients with chronic periodontitis, which represents a frequency of 34.5%. All 30 isolates (100%) were sensitive to metronidazole, with MIC values ranging from 0015-4ug/ml. Regarding tetracycline, 27 isolates (90%) were sensitive, with MIC values ranging from <0.015 to 4 ug /ml, the remaining three isolates (10%) were resistant to tetracycline with MIC values of 8ug/ ml. There was no statistically significant difference in age, gender, pocket depth, clinical attachment level and severity of periodontitis between the group of patients with chronic periodontitis and P. gingivalis and the group of patients with chronic periodontitis without P. gingivalis. In conclusion, P. gingivalis was found at a frequency of 34.5% in patients with chronic periodontitis and clinical isolates were highly sensitive to metronidazole and tetracycline.

Occurrence of porphyromonas gingivalis and its antibacterial susceptibility to metronidazole and tetracycline in patients with chronic periodontitis. / Occurrence of porphyromonas gingivalis and its antibacterial susceptibility to metronidazole and tetracycline in patients with chronic periodontitis.

Chronic periodontitis is a multifactorial infectious disease associated with Gram-negative strict anaerobes which are immersed in the subgingival biofilm. Porphyromonas gingivalis, an important periodontal pathogen, is frequently detected in patients with chronic periodontitis. Although isolates of P. gingivalis tend to be susceptible to most antimicrobial agents, relatively little information is available on its in vitro antimicrobial susceptibility. The aim of this study was to determine the frequency of P. gingivalis in patients with chronic periodontitis and to assess antimicrobial susceptibility in terms of minimum inhibitory concentration (MIC) of clinical isolates to metronidazole and tetracycline. A descriptive, observational study was performed including 87 patients with chronic periodontitis. Samples were taken from the periodontal pocket using paper points, which were placed in thioglycollate broth. Samples were incubated for 4 hours at 37°C in anaerobic conditions and finally replated on Wilkins-Chalgren anaerobic agar (Oxoid). Bacteria were identified using the RapIDTMANAII system (Remel) and antimicrobial susceptibility was determined with the M.I.C. Evaluator test (MICE, Oxoid). P. gingivalis was identified in 30 of the 87 patients with chronic periodontitis, which represents a frequency of 34.5

. All 30 isolates (100

) were sensitive to metronidazole, with MIC values ranging from 0015-4ug/ml. Regarding tetracycline, 27 isolates (90

) were sensitive, with MIC values ranging from <0.015 to 4 ug /ml, the remaining three isolates (10

) were resistant to tetracycline with MIC values of 8ug/ ml. There was no statistically significant difference in age, gender, pocket depth, clinical attachment level and severity of periodontitis between the group of patients with chronic periodontitis and P. gingivalis and the group of patients with chronic periodontitis without P. gingivalis. In conclusion, P. gingivalis was found at a frequency of 34.5

in patients with chronic periodontitis and clinical isolates were highly sensitive to metronidazole and tetracycline.

Presence and antimicrobial profile of gram-negative facultative anaerobe rods in patients with chronic periodontitis and gingivitis.

Chronic periodontitis is a multifactorial infectious disease associated with Gram-negative anaerobes which are part of the subgingival microflora. In recent years, studies have been conducted to assess the presence of Gram-negative facultative anaerobes (Enterobacteriaceae) and their participation in the development and progression of chronic periodontitis. The aim of this study was to determine the presence of Enterobacteriaceae in patients with chronic periodontitis and gingivitis and to assess antimicrobial susceptibility of clinical isolates. A descriptive, observational study was performed including 64 patients with chronic periodontitis and 22 patients with gingivitis. Microbiological samples were taken from the gingival sulcus using paper points, which then were placed in thioglycollate broth. Samples were incubated for 4 hours at 37 degrees C and finally replated on MacConkey agar Bacteria were identified using the API-20E system (Biomerieux, France) and antimicrobial susceptibility was determined using the disk diffusion method. The evaluation of samples showed presence of 29 enterobacterial species distributed as follows: 7 in the group with gingivitis and 22 in the group with chronic periodontitis. In the chronic periodontitis group the most common species were: K. oxytoca n = 5, S. liquefaciens n = 4 and K. pneumoniae and E. coli with n = 3. The gingivitis group had the highest frequency of Erwinia sp. (n = 2). Clinical isolates showed very low sensitivity levels to beta-lactam ampicillin and amoxicillin/ clavulanic acid, 17.2% and 27.6% respectively, and higher sensitivity levels to ciprofloxacin (96.6%), amikacin (79.3%), gentamicin (68.9%) and ceftazidime, ceftriaxone, kanamycin and trimethoprimsulfa (65.5%). In conclusion, the existence of a high frequency of enterobacteria in patients with chronic periodontitis and gingivitis shows that periodontologists should pay greater attention to prevention protocols, and develop mechanical and antimicrobial therapies in which antimicrobial susceptibility profile reports should be considered as part of periodontal treatment.

Antimicrobial activity of fractions and subfractions of Elaeagia utilis against microorganisms of importance in dental caries.

Dental caries is a multifactorial infectious disease that leads to the destruction of dental hard tissue. The main goal of research into medicinal plants is to seek compounds with antimicrobial activity for subsequent use in prevention strategies and control of infectious diseases. The aim of this study was to evaluate the antimicrobial activity of fractions and subfractions obtained from Elaeagia utilis against Streptococcus mutans, Streptococcus sobrinus and Lactobacillus acidophilus. The plant material was collected in the town of Alban (Cundinamarca, Colombia), which is located at an altitude of 2245 meters above sea level. Two extracts were obtained by cold maceration of E. utilis leaves in (a) petroleum ether extract and (b) ethanol extract. Fractions were obtained from the petroleum ether extract by column vacuum chromatography, and from the ethanol extract by continuous liquid/liquid partitioning. The antimicrobial activity of fractions and subfractions was evaluated by the well diffusion method. At a concentration of 10 mg/well, several fractions from both extracts showed antimicrobial activity against S. mutans, S. sobrinus and L. acidophilus. Among the ethanol extract fractions, the dichloromethane fraction had notably greater antimicrobial activity. It was sub-partitioned, yielding three subfractions with inhibitory activity, of which the most active was MeOH: H2O (Bp) with minimum inhibitory concentration 0.1 mg/well on the 3 study bacteria. Terpenes, sesquiterpenlactones and simple phenolic compounds were identified in it. In conclusion, this study shows the antimicrobial potential of fractions and subfractions obtained from extracts of E. utilis leaves against bacteria that are important in dental caries.

Presence and antimicrobial profile of Gram-negative facultative anaerobe rods in patients with chronic periodontitis and gingivitis

La periodontitis crónica es una enfermedad infecciosa multifactorial hacen parte de la microflora subgingival. En los últimos años se han realizado estudios para valorar la presencia de bacilos Gramnegativos anaerobios facultativos (enterobacterias) y suimportancia en el desarrollo y rogresión de la periodontitis crónica. El objetivo de este estudio fue determinar la presencia de enterobacterias en pacientes con periodontitis crónica y gingivitis y conocer la susceptibilidad antimicrobiana de los aislamientosclínicos. Se realizó un estudio observacional y descriptivo en elque se incluyeron 64 pacientes con periodontitis crónica y 22 pacientes con gingivitis. Las muestras tomadas en el surco gingival con conos de papel se depositaron en caldo tioglicolato, seincubaron durante 4 horas a 37 oC y se resembraron finalmente en Agar MacConkey. En la identificación de las bacterias se utilizó el sistema API-20E (Biomerieux, France) y la susceptibilidadantimicrobiana se realizó por el método de difusión en disco. En los dos grupos se identificaron 29 especies enterobacterianas, 7 en el grupo con gingivitis y 22 en el grupo con periodontitis crónica. En el grupo de periodontitis crónica las especies masfrecuentes fueron: K. oxytoca n=5, S. liquefaciens n=4 y K.pneumoniaey E. coli con n=3. En el grupo con gingivitis, Erwiniasp tuvo la mayor frecuencia (n=2). Los aislamientos clínicos presentaron níveles muy bajos de sensibilidad a los B-lactamicosampicilina y amoxicilina/ ac.clavulanico, 17.2 y 27.6 por ciento, y la mayor sensibilidad a ciprofloxacina. En conclusión, la alta frecuencia de enterobacterias en pacientes con periodontitis debe conducir a la prevención y a desarrollar terapias mecánicas y antimicrobianas en las cuales se tengan en cuenta, como parte del tratamiento periodontal, los perfiles antimicrobianos reportados.

Antimicrobial activity of fractions and subfractions of Elaeagia utilis against microorganisms of importance in dental caries

La caries dental es una enfermedad infecciosa multifactorial que conduce a la destrucción del tejido duro dental. El principal objetivo de la investigación en plantas medicinales es la búsquedade compuestos con actividad antimicrobiana para su posterior uso en estrategias de prevención o control de enfermedades infecciosas. El objetivo de este estudio fue evaluar la actividad antimicrobiana de fracciones y subfracciones obtenidas de la planta Elaeagia utilis contra Streptococcus mutans, Streptococcus sobrinus y Lactobacillus acidophilus. El materialvegetal fue colectado en la ciudad de Albán (Cundinamarca-Colombia) situada a una altitud de 2245 metros sobre el nivel del mar. Mediante el método de maceración en frio de hojas de E. utilis se obtuvieron dos extractos, uno en éter de petróleo y otro en etanol. Del extracto etéreo se obtuvieron fraccionesmediante cromatografía en columna al vacío y al extracto etanólico se le realizó fraccionamiento líquido/líquido continuo. La evaluación de la actividad antimicrobiana de las fracciones ysubfracciones se realizó por el método de difusión en pozo. A una concentración de 10 mg/pozo, múltiples fracciones obtenidas de los dos extractos presentaron actividad antimicrobianasobre S. mutans, S. sobrinus y L. acidophilus. De las fracciones del extracto etanólico se destaca la fracción diclorometano, por presentar mayor actividad antimicrobiana, razón por lo cual se subfracciona y se obtienen tres subfracciones con actividad inhibitoria.La subfracción más activa fue eOH:H2O (Bp) con una concentración mínima inhibitoria de 0.1 mg/pozo sobre las 3bacterias en estudio. En esta subfracción se determinaron terpenos, sesquiterpenlactonas y compuestos fenólicos simples. En conclusión, en este estudio se presenta el potencial antimicrobiano de fracciones y subfracciones obtenidas de extractos dehojas de E. utilis contra microorganismos de importancia encaries dental.

Dental caries is a multifactorial infectious disease that leads tothe destruction of dental hard tissue. The main goal of researchinto medicinal plants is to seek compounds with antimicrobialactivity for subsequent use in prevention strategies and controlof infectious diseases. The aim of this study was to evaluate theantimicrobial activity of fractions and subfractions obtainedfrom Elaeagia utilis against Streptococcus mutans, Streptococ-cus sobrinus and Lactobacillus acidophilus. The plant materialwas collected in the town of Alban (Cundinamarca, Colombia),which is located at an altitude of 2245 meters above sea level.Two extracts were obtained by cold maceration of E. utilis leavesin (a) petroleum ether extract and (b) ethanol extract. Fractionswere obtained from the petroleum ether extract by column vacu-um chromatography, and from the ethanol extract by continuousliquid / liquid partitioning. The antimicrobial activity of frac-tions and subfractions was evaluated by the well diffusionmethod. At a concentration of 10 mg /well, several fractions fromboth extracts showed antimicrobial activity against S. mutans, S.sobrinus and L. acidophilus. Among the ethanol extract frac-tions, the dichloromethane fraction had notably greaterantimicrobial activity. It was sub-partitioned, yielding three sub-fractions with inhibitory activity, of which the most active wasMeOH: H2O (Bp) with minimum inhibitory concentration 0.1mg /well on the 3 study bacteria. Terpenes, sesquiterpenlactonesand simple phenolic compounds were identified in it. In conclu-sion, this study shows the antimicrobial potential of fractionsand subfractions obtained from extracts of E. utilis leaves againstbacteria that are important in dental caries.

Presence and antimicrobial profile of gram-negative facultative anaerobe rods in patients with chronic periodontitis and gingivitis. / Presence and antimicrobial profile of gram-negative facultative anaerobe rods in patients with chronic periodontitis and gingivitis.

Chronic periodontitis is a multifactorial infectious disease associated with Gram-negative anaerobes which are part of the subgingival microflora. In recent years, studies have been conducted to assess the presence of Gram-negative facultative anaerobes (Enterobacteriaceae) and their participation in the development and progression of chronic periodontitis. The aim of this study was to determine the presence of Enterobacteriaceae in patients with chronic periodontitis and gingivitis and to assess antimicrobial susceptibility of clinical isolates. A descriptive, observational study was performed including 64 patients with chronic periodontitis and 22 patients with gingivitis. Microbiological samples were taken from the gingival sulcus using paper points, which then were placed in thioglycollate broth. Samples were incubated for 4 hours at 37 degrees C and finally replated on MacConkey agar Bacteria were identified using the API-20E system (Biomerieux, France) and antimicrobial susceptibility was determined using the disk diffusion method. The evaluation of samples showed presence of 29 enterobacterial species distributed as follows: 7 in the group with gingivitis and 22 in the group with chronic periodontitis. In the chronic periodontitis group the most common species were: K. oxytoca n = 5, S. liquefaciens n = 4 and K. pneumoniae and E. coli with n = 3. The gingivitis group had the highest frequency of Erwinia sp. (n = 2). Clinical isolates showed very low sensitivity levels to beta-lactam ampicillin and amoxicillin/ clavulanic acid, 17.2

and 27.6

respectively, and higher sensitivity levels to ciprofloxacin (96.6

), amikacin (79.3

), gentamicin (68.9

) and ceftazidime, ceftriaxone, kanamycin and trimethoprimsulfa (65.5

). In conclusion, the existence of a high frequency of enterobacteria in patients with chronic periodontitis and gingivitis shows that periodontologists should pay greater attention to prevention protocols, and develop mechanical and antimicrobial therapies in which antimicrobial susceptibility profile reports should be considered as part of periodontal treatment.

Antimicrobial activity of fractions and subfractions of Elaeagia utilis against microorganisms of importance in dental caries. / Antimicrobial activity of fractions and subfractions of Elaeagia utilis against microorganisms of importance in dental caries.

Dental caries is a multifactorial infectious disease that leads to the destruction of dental hard tissue. The main goal of research into medicinal plants is to seek compounds with antimicrobial activity for subsequent use in prevention strategies and control of infectious diseases. The aim of this study was to evaluate the antimicrobial activity of fractions and subfractions obtained from Elaeagia utilis against Streptococcus mutans, Streptococcus sobrinus and Lactobacillus acidophilus. The plant material was collected in the town of Alban (Cundinamarca, Colombia), which is located at an altitude of 2245 meters above sea level. Two extracts were obtained by cold maceration of E. utilis leaves in (a) petroleum ether extract and (b) ethanol extract. Fractions were obtained from the petroleum ether extract by column vacuum chromatography, and from the ethanol extract by continuous liquid/liquid partitioning. The antimicrobial activity of fractions and subfractions was evaluated by the well diffusion method. At a concentration of 10 mg/well, several fractions from both extracts showed antimicrobial activity against S. mutans, S. sobrinus and L. acidophilus. Among the ethanol extract fractions, the dichloromethane fraction had notably greater antimicrobial activity. It was sub-partitioned, yielding three subfractions with inhibitory activity, of which the most active was MeOH: H2O (Bp) with minimum inhibitory concentration 0.1 mg/well on the 3 study bacteria. Terpenes, sesquiterpenlactones and simple phenolic compounds were identified in it. In conclusion, this study shows the antimicrobial potential of fractions and subfractions obtained from extracts of E. utilis leaves against bacteria that are important in dental caries.

Presence and antimicrobial profile of Gram-negative facultative anaerobe rods in patients with chronic periodontitis and gingivitis

La periodontitis crónica es una enfermedad infecciosa multifactorial hacen parte de la microflora subgingival. En los últimos años se han realizado estudios para valorar la presencia de bacilos Gramnegativos anaerobios facultativos (enterobacterias) y suimportancia en el desarrollo y rogresión de la periodontitis crónica. El objetivo de este estudio fue determinar la presencia de enterobacterias en pacientes con periodontitis crónica y gingivitis y conocer la susceptibilidad antimicrobiana de los aislamientosclínicos. Se realizó un estudio observacional y descriptivo en elque se incluyeron 64 pacientes con periodontitis crónica y 22 pacientes con gingivitis. Las muestras tomadas en el surco gingival con conos de papel se depositaron en caldo tioglicolato, seincubaron durante 4 horas a 37 oC y se resembraron finalmente en Agar MacConkey. En la identificación de las bacterias se utilizó el sistema API-20E (Biomerieux, France) y la susceptibilidadantimicrobiana se realizó por el método de difusión en disco. En los dos grupos se identificaron 29 especies enterobacterianas, 7 en el grupo con gingivitis y 22 en el grupo con periodontitis crónica. En el grupo de periodontitis crónica las especies masfrecuentes fueron: K. oxytoca n=5, S. liquefaciens n=4 y K.pneumoniaey E. coli con n=3. En el grupo con gingivitis, Erwiniasp tuvo la mayor frecuencia (n=2). Los aislamientos clínicos presentaron níveles muy bajos de sensibilidad a los B-lactamicosampicilina y amoxicilina/ ac.clavulanico, 17.2 y 27.6 por ciento, y la mayor sensibilidad a ciprofloxacina. En conclusión, la alta frecuencia de enterobacterias en pacientes con periodontitis debe conducir a la prevención y a desarrollar terapias mecánicas y antimicrobianas en las cuales se tengan en cuenta, como parte del tratamiento periodontal, los perfiles antimicrobianos reportados.(AU)

Antimicrobial activity of fractions and subfractions of Elaeagia utilis against microorganisms of importance in dental caries

La caries dental es una enfermedad infecciosa multifactorial que conduce a la destrucción del tejido duro dental. El principal objetivo de la investigación en plantas medicinales es la búsquedade compuestos con actividad antimicrobiana para su posterior uso en estrategias de prevención o control de enfermedades infecciosas. El objetivo de este estudio fue evaluar la actividad antimicrobiana de fracciones y subfracciones obtenidas de la planta Elaeagia utilis contra Streptococcus mutans, Streptococcus sobrinus y Lactobacillus acidophilus. El materialvegetal fue colectado en la ciudad de Albán (Cundinamarca-Colombia) situada a una altitud de 2245 metros sobre el nivel del mar. Mediante el método de maceración en frio de hojas de E. utilis se obtuvieron dos extractos, uno en éter de petróleo y otro en etanol. Del extracto etéreo se obtuvieron fraccionesmediante cromatografía en columna al vacío y al extracto etanólico se le realizó fraccionamiento líquido/líquido continuo. La evaluación de la actividad antimicrobiana de las fracciones ysubfracciones se realizó por el método de difusión en pozo. A una concentración de 10 mg/pozo, múltiples fracciones obtenidas de los dos extractos presentaron actividad antimicrobianasobre S. mutans, S. sobrinus y L. acidophilus. De las fracciones del extracto etanólico se destaca la fracción diclorometano, por presentar mayor actividad antimicrobiana, razón por lo cual se subfracciona y se obtienen tres subfracciones con actividad inhibitoria.La subfracción más activa fue eOH:H2O (Bp) con una concentración mínima inhibitoria de 0.1 mg/pozo sobre las 3bacterias en estudio. En esta subfracción se determinaron terpenos, sesquiterpenlactonas y compuestos fenólicos simples. En conclusión, en este estudio se presenta el potencial antimicrobiano de fracciones y subfracciones obtenidas de extractos dehojas de E. utilis contra microorganismos de importancia encaries dental.(AU)

Dental caries is a multifactorial infectious disease that leads tothe destruction of dental hard tissue. The main goal of researchinto medicinal plants is to seek compounds with antimicrobialactivity for subsequent use in prevention strategies and controlof infectious diseases. The aim of this study was to evaluate theantimicrobial activity of fractions and subfractions obtainedfrom Elaeagia utilis against Streptococcus mutans, Streptococ-cus sobrinus and Lactobacillus acidophilus. The plant materialwas collected in the town of Alban (Cundinamarca, Colombia),which is located at an altitude of 2245 meters above sea level.Two extracts were obtained by cold maceration of E. utilis leavesin (a) petroleum ether extract and (b) ethanol extract. Fractionswere obtained from the petroleum ether extract by column vacu-um chromatography, and from the ethanol extract by continuousliquid / liquid partitioning. The antimicrobial activity of frac-tions and subfractions was evaluated by the well diffusionmethod. At a concentration of 10 mg /well, several fractions fromboth extracts showed antimicrobial activity against S. mutans, S.sobrinus and L. acidophilus. Among the ethanol extract frac-tions, the dichloromethane fraction had notably greaterantimicrobial activity. It was sub-partitioned, yielding three sub-fractions with inhibitory activity, of which the most active wasMeOH: H2O (Bp) with minimum inhibitory concentration 0.1mg /well on the 3 study bacteria. Terpenes, sesquiterpenlactonesand simple phenolic compounds were identified in it. In conclu-sion, this study shows the antimicrobial potential of fractionsand subfractions obtained from extracts of E. utilis leaves againstbacteria that are important in dental caries.(AU)

Antimicrobial potential of extracts from Stevia rebaudiana leaves against bacteria of importance in dental caries.

In recent years, the antimicrobial activity of Stevia rebaudiana Bertoni leaf extracts against a large number of microorganisms has been evaluated, but not its activity against microorganisms of importance in dental caries. The aim of this study was to evaluate the antibacterial activity of Stevia rebaudiana Bertoni leaf extracts against cariogenic bacteria. Extracts were obtained from the dried Stevia rebaudiana Bertoni leaves in hexane, methanol, ethanol, ethyl acetate and chloroform. The antimicrobial activity of the 5 extracts against 16 bacterial strains of the genera Streptococcus (n= 12) and Lactobacillus (n= 4) was evaluated by the well diffusion method. Minimal inhibitory concentrations (MIC) of the extracts in hexane, methanol, ethanol, ethyl acetate and chloroform on the 16 bacterial strains were respectively 30 mg/ml, 120 mg/ml, 120 mg/ml, 60 mg/ml and 60 mg/ml. The zones of inhibition present at the MIC were variable, ranging from 9 mm to 17.3 mm. Our results suggest that inhibition zones with a hexane extract are similar to those obtained with ethanol and methanol, but the minimal inhibitory concentration (30 mg/ml) is lower. For the four Lactobacillus species, the inhibition zones obtained between 12.3 and 17.3 mm were somewhat larger with ethyl acetate and chloroform extracts, suggesting they were the most susceptible microorganisms.

Antimicrobial potential of extracts from Stevia Rebaudiana leaves against bacteria of importance in dental caries

En los últimos años, se ha evaluado la actividad antimicrobiana de extractos obtenidos de jojas de Stevia rebaudiana Bertoni sobre un gran número de microorganismos de importancia encaries dental. El objetivo de este estudio fue evaluar la actividad antibacteriana de extractos de hojas de Stevia rebaudiana Bertoni sobre microorganismos cariogénicos. A partir de hojas secas de Stevia rebaudiana Bertoni convertidas en polvillo seobtuvieron los extractos en dichos solventes. La evaluación de la actividad antimicrobiana de los 5 extractos sobre las 16 cepas bacterianas de los géneros Streptococcus (n=12) y Lactobacillus(n=4) se realizó por el método de difusión en pozo. Las concentraciones mínimas inhibitorias (CMI) de los extractos de hexano, metanol, etanol, acetato de etilo y cloroformo, sobre las 16 cepas bacterianas fueron respectivamente de 30 mg/ml, 120 mg/ml, 120 mg/ml, 60 mg/ml y 60 mg/ml, respectivamente. Los halos de inhibición determinados a las CMIfueron variables, el de menor valor fue 9 mm y el de mayor fue de 17.3 mm. Nuestros resultados, sugieren que los halos de inhibición en el extracto de hexano son semejantes a los obtenidos para el etanol y metanol, sin embargo, la CMI (30 mg/ml) es menor. En las 4 especies de Lactobacillus los halosde inhibición obtenidos entre 13 y 17.3 mm, son ligeramente mayores en los extractos de acetato de etilo y cloroformo, sugiriendo que fueron los microorganismos más susceptibles.

Antimicrobial potential of extracts from Stevia Rebaudiana leaves against bacteria of importance in dental caries

En los últimos años, se ha evaluado la actividad antimicrobiana de extractos obtenidos de jojas de Stevia rebaudiana Bertoni sobre un gran número de microorganismos de importancia encaries dental. El objetivo de este estudio fue evaluar la actividad antibacteriana de extractos de hojas de Stevia rebaudiana Bertoni sobre microorganismos cariogénicos. A partir de hojas secas de Stevia rebaudiana Bertoni convertidas en polvillo seobtuvieron los extractos en dichos solventes. La evaluación de la actividad antimicrobiana de los 5 extractos sobre las 16 cepas bacterianas de los géneros Streptococcus (n=12) y Lactobacillus(n=4) se realizó por el método de difusión en pozo. Las concentraciones mínimas inhibitorias (CMI) de los extractos de hexano, metanol, etanol, acetato de etilo y cloroformo, sobre las 16 cepas bacterianas fueron respectivamente de 30 mg/ml, 120 mg/ml, 120 mg/ml, 60 mg/ml y 60 mg/ml, respectivamente. Los halos de inhibición determinados a las CMIfueron variables, el de menor valor fue 9 mm y el de mayor fue de 17.3 mm. Nuestros resultados, sugieren que los halos de inhibición en el extracto de hexano son semejantes a los obtenidos para el etanol y metanol, sin embargo, la CMI (30 mg/ml) es menor. En las 4 especies de Lactobacillus los halosde inhibición obtenidos entre 13 y 17.3 mm, son ligeramente mayores en los extractos de acetato de etilo y cloroformo, sugiriendo que fueron los microorganismos más susceptibles.(AU)