Comparative evaluation of antimicrobial potential of herbal extracts on Streptococcus mutans and Enterococcus faecalis An in vitro study / Avaliação comparativa do potencial antimicrobiano de extratos de ervas em Streptococcus mutans e Enterococcus faecalis: estudo in vitro

Though aloe vera extract, green tea extract and coriander oil are proven antimicrobial agents, very little information is available regarding its effects on oral bacteria, Streptococcus mutans, which is responsible for initiating caries and Enterococcus faecalis, responsible for failure of root canal treatment. Objective: To find the antimicrobial activity of aloe vera extract, black tea extract and coriander oil against S. mutans and E. faecalis. Materials and Methods: The agar well diffusion method was used to determine the antibacterial activity of Aloe vera extract, black tea extract and coriander oil. Different concentration of prepared plant extracts and coriander seed oil (50 & 100 µl) was incorporated into the wells and the plates containing S. mutans and E. faecalis were incubated at 37 °C for 24 h. The antibiotic (amoxicillin 30 µl) was used as positive control. Zone Of Inhibition (ZOI) was recorded in each plate. Results: For S. mutans, the maximum ZOI was created by coriander oil with a diameter of 25.00±0.58 mm at 50 µl and for E. faecalis, maximum ZOI was created by aloe vera extract 16.00±0.58 mm at 100 µl concentration which were far better than the control: amoxicillin 30 µl concentration. Conclusion: The extracts of Aloe vera, black tea and coriander oil, showed significant activity against the investigated microbial strains, Streptococcus mutans and Enterococcus faecalis which further helps in the development of new topical agents that help in reducing the numbers of these organisms present in the oral cavity. (AU)

Embora o extrato de aloe vera, extrato de chá verde e óleo de coentro sejam agentes antimicrobianos comprovados, há pouca informação disponível sobre seus efeitos nas bactérias orais, Streptococcus mutans, que é responsável por iniciar cáries e Enterococcus faecalis, responsável pela falha do tratamento de canal radicular. Objetivo: Avaliar a atividade antimicrobiana do extrato de aloe vera, extrato de chá preto e óleo de coentro contra S. mutans e E. faecalis. Materiais e Métodos: O método de difusão em agar foi usado para determinar a atividade antibacteriana do extrato de Aloe vera, extrato de chá preto e óleo de coentro. Diferentes concentrações dos extratos de plantas e óleo de semente de coentro (50 e 100 µl) foram preparados e colocados nos poços e nas placas contendo S. mutans e E. faecalis e foram incubadas a 37°C por 24 h. O antibiótico (amoxicilina 30 µl) foi utilizado como controle positivo. A zona de inibição (ZOI) foi registrada em cada placa. Resultados: Para S. mutans, a ZOI máxima foi obtida com o óleo de coentro com um diâmetro de 25,00 ± 0,58 mm a 50 µl e para E. faecalis, a ZOI máxima foi obtiada pelo extrato de aloe vera 16,00 ± 0,58 mm na concentração de 100 µl, as quais foram melhores do que o controle: concentração de 30 µl de amoxicilina. Conclusão: Os extratos de Aloe vera, chá preto e óleo de coentro apresentaram atividade significativa contra as cepas microbianas investigadas, Streptococcus mutans e Enterococcus faecalis auxiliando no desenvolvimento de novos agentes tópicos visando a redução do número desses organismos presentes no cavidade oral. (AU)

Antibiotic resistance of Enterococcus faecalis from persistent endodontic infections / Resistencia antibiótica de Enterococcus faecalis provenientes de infecciones endodónticas persistentes

Enterococci are important nosocomial pathogens due to their intrinsic multiresistance and the acquisition of new antibiotic resistance genes (ARG). Enterococcus faecalis has been shown to be one of the main pathogens in persistent endodontic infections, therefore, the main objective of this study was to evaluate the phenotype and resistance genotype of strains of E. faecalis isolated from teeth with persistent endodontic lesions, to the most commonly prescribed antibiotics in dentistry. Thirteen strains of E. faecalis of different pulsotype were analyzed to evaluate the susceptibility to antibiotics, amoxicillin, amoxicillin/clavulanic acid, tetracycline, erythromycin and metronidazole, using the Epsilometer test (E- test) and the presence of beta-lactamases with nitrocefin test. Finally, the detection of ARG was performed with a molecular polymerase chain reaction (PCR) technique and confirmed by the sequencing of the amplification products. Fisher's exact test was used, using 95 % confidence. Regarding the phenotype of resistance, the evaluated strains, independent of the pulsotype, were totally resistant to the action of metronidazole. Antibiotics with higher minimum inhibitory concentration (MIC) after metronidazole include tetracycline and erythromycin. In contrast, lower MIC are applied to the combination of amoxicillin with clavulanic acid. The nitrocefin test was positive only in one strain. Genotypically, two genetically distant strains isolated from a single patient, presented a genotype of resistance to erythromycin, determined by the presence of the ermB gene. No statistically significant relationship was found between phenotypic resistance and the presence of ARG in relation to erythromycin (p> 0.05). It was concluded that isolates of E. faecalis from persistent endodontic infections showed phenotypes of resistance to several antimicrobial agents, all of which were susceptible to amoxicillin/clavulanic acid. Periodic evaluation of susceptibility to antibiotics is suggested as an important practice for the surveillance of antibiotic resistance in oral strains.

Los enterococos son importantes patógenos nosocomiales debido a su multi resistencia intrínseca y la adquisición de nuevos genes de resistencia a los antibióticos (ARG). Enterococcus faecalis es uno de los principales patógenos en infecciones endodónticas persistentes, por lo tanto, el objetivo principal de este estudio fue evaluar el fenotipo y el genotipo de resistencia de cepas de E. faecalis aisladas de dientes con lesiones endodóncicas persistentes, a los antibióticos comúnmente recetados en odontología. Se analizaron 13 cepas de E. faecalis de diferentes pulsotipos para evaluar la susceptibilidad a los antibióticos, amoxicilina, amoxicilina / ácido clavulánico, tetraciclina, eritromicina y metronidazol, utilizando la prueba de Epsilometría (E-test) y la presencia de beta-lactamasas con prueba de nitrocefina. Finalmente, la detección de ARG se realizó con una técnica molecular de reacción en cadena de la polimerasa (PCR) y se confirmó mediante la secuenciación de los productos de amplificación. Se utilizó la prueba exacta de Fisher, con un 95 % de confianza. En cuanto al fenotipo de resistencia, las cepas evaluadas, independientes del pulsotipo, fueron totalmente resistentes a la acción del metronidazol. Los antibióticos con los valores más altos de concentración mínima inibitoria (CMI) después del metronidazol incluyen tetraciclina y eritromicina. En contraste, las CMI mas bajas se aplican a la combinación de amoxicilina con ácido clavulánico. La prueba de nitrocefina fue positiva solo en una cepa. Genotípicamente, dos cepas distantes genéticamente, aisladas de un mismo paciente fueron positivas para el gen ermB. No se encontró una relación estadísticamente significativa entre la resistencia fenotípica y la presencia de ARG en relación con la eritromicina (p> 0,05). Se concluyó que los aislamientos de E. faecalis de infecciones endodónticas persistentes mostraron fenotipos de resistencia a varios agentes antimicrobianos, todos los cuales fueron susceptibles a amoxicilina / ácido clavulánico. Se sugiere una evaluación periódica de la susceptibilidad a los antibióticos como una práctica importante para la vigilancia de la resistencia a los antibióticos en las cepas orales.

Comparación de la eficacia de los irrigantes OxOral ® y NaOCl en la eliminación de Enterococcus faecalis / Comparison of OxOral ® and NaOCl irrigants efficiency in Enterococcus faecalis elimination

Resumen Objetivo: Comparar la eficacia en la eliminación de Enterococcus faecalis con OxOral® versus hipoclorito de sodio a los 15 y 60 segundos. Material y métodos: Se incluyeron 36 cultivos de E. faecalis ATCC 29212 asignados en dos grupos; OxOral® e hipoclorito de sodio al 5.25% que a su vez fueron divididos en 15 y 60 segundos. Se colocaron 8 mL de agua peptonada, 1 mL del irrigante y 1 mL de la cepa, se dejó reposar. A cada tiempo se extrajo 1 mL y se sembró en agar sangre por 24 horas. Se empleó и de Mann-Whitney. Resultados: Con hipoclorito de sodio a 15 segundos hubo tres cultivos con crecimiento aceptable y seis extendido; a los 60 segúndos, cuatro tuvieron resultado eficaz, tres aceptable, uno extendido. Con OxOral® hubo crecimiento extendido en los nueve cultivos, en ambos tiempos, encontrando diferencias estadísticamente significativas a los 60 segundos (p < 0.01). Conclusión: La eliminación de E. faecalis fue mejor con hipoclorito de sodio a los 60 segundos.

ABSTRACT Objective: To compare effectiveness of ОхОrаl® versus sodium hypochlorite in Enterococcus faecalis elimination at 15 and 60 seconds. Material and methods: Material used in the study was 36 E, faecalis ATCC 29212 cultures assigned to two groups: ОхОrаl® and 5.25% sodium hypochlorite. Both groups were in turn divided into 15 and 60 second samples. Samples were placed in peptone water, 1 mL of irrigating solution and 1 mL of strain were left to rest. 1 mL was extracted at each time, samples were seeded into blood agar for 24 hours. Mann-Whitney и test was applied. Results: With sodium hypochlorite at 15 seconds, there were three cultures with acceptable growth and six with extended growth; at 60 seconds four cultures exhibited effective result, three acceptable and one extended. With ОхОrаl® there was extended growth in all nine cultures at both established times, significant statistical differences were found at the 60 seconds time (p < 0.01). Conclusion: E, faecalis elimination was better with sodium hypochlorite at 60 seconds.

Role of TRAF6 in inflammatory responses of human osteoblast-like cells with Enterococcusfaecalis / 口腔疾病防治

Objective @#Explore the role and status of tumor necrosis factor receptor-associated factor 6 (TRAF6) in the inflammatory response of human osteoblast-like cells MG63 which was triggered by Enterococcus faecalis (E. faecalis) and its lipoteichoic acid (LTA)@*Methods@# SiRNA technology was applied to silence the TRAF6 gene of MG63 cells, Using E.faecelis and its LTA to stimulate the silence MG63 cells with different hours. After that, using real-time PCR technology to detect toll-like receptor 2 (TLR2) and TRAF6 gene expression and using ELISA assay to detect proinflammatory cytokines interleukin-1β and TNF-alpha expression levels.@*Results@#When MG63 cells was infected by E. faecalis, its LTA, TLR2 and TRAF6 gene level has increased to varying degrees (P< 0.05); interleukin-1β and TNF-alpha expression was significantly higher (P< 0.05). When TRAF6 gene of MG63 cells was silenced by siRNA, pro-inflammatory cytokines interleukin-1β, interleukin-6, interleukin -8 and TNF-alpha expression decreased significantly (P< 0.05).@*Conclusion@#E. faecalis and its toxic components is identified by MG63 cells mainly through TLR2 receptors. The major virulence factor in periapical infections caused by E. faecalis is LTA.

Bactericidal effect of nano-silver against E.faecalis growing in multi-species biofilm / 实用口腔医学杂志

Objective:To evaluate the bactericidal effects of nano-silver against Enterococcus faecalis(E.faecalis) growing in multi-species biofilm.Methods:85 biofilms were established using MBECTM P&G Assay with E.faecalis together with Fusobacterium nucleatum and P.melaninogenica.Thereafter,10 specimens were used as the controls,75 were randomly divided into 3 groups(n=25),and treated with 0.1% nano-silver (12-15 nm) solution,0.1% nano-silver (100 nm) solution and 2% hypochlorite solution,respectively.Each sample was then separated into 2 different tubes.PMA was added to one of the tubes,and the other was left untreated.Then,DNA extraction and qPCR were performed.The cycle threshold(Ct) values between samples were compared by paired t test.Results:The Ct values of the samples treated with PMA were higher than that without PMA(P=0.000) in the group of 0.1% nano-silver solution(12-15 nm)was higher than that in the group of 0.1% nano-silver solution(100 nm)and 2% sodium hypochlorite solution.(P<0.05);the value in the group of 0.1% nano-silver solution(100 nm)was larger than that in 2% sodium hypochlorite solution(P<0.05).Conclusion:0.1% nano silver solution might have a strong bactericidal effect against against E.faecalis growing in multi-species biofilm.The bactericidal effect may be enhanced with the small size of silver particles.

Comparative Evaluation Of Anti – Bacterial Efficacy Of Sodium Hypochlorite And Carbolic Acid Against Enterococcus Faecalis As Root Canal Irrigant.

Background: Elimination of microorganisms from infected root canal is a complicated & difficult task. Elimination or sufficient reduction of irritants, microorganisms and prevention of recontamination of the root canal after the treatment is the essential factor for successes of the treatment. Methods: Following materials required for this study i.e. 36 teeth prepared for root canal inoculation, Enterococcus faecalis strain, 5 % and 10 % carbolic acid solution, 5.25 % sodium hypochlorite solution, phosphate buffer saline and normal saline solution. An overnight subculture plate of E.faecalis was taken for inoculum preparation and uniform suspension made in normal saline (02 ml) by taking up isolated colonies and adjusted to 0.5 McFarland standard which gives 1.5 X 108 cfu / ml. Result and Conclusion: From this study, we can conclude that 5.25 % NaOCl is one of the superior and successful endodontic irrigant against 5 % and 10% carbolic acid.In our study, result shows that sodium hypochlorite solution is superior to carbolic acid solution. However, as this study is in vitro study, not simulate the oral environment; further few clinical studies are required in evaluation of antimicrobial efficacy of these solutions for predictable and successful endodontic outcome.

Quantitative evaluation of live Enterococcus faecalis in failed endodontic root-well-filled teeth / 实用口腔医学杂志

Objective:To analyse viable Enterococcus faecalis(E.faecalis)in root-well-filled teeth associated with failed endodontic treatment by using propidium monoazide (PMA)in combination with real time qPCR.Methods:Bacterial samples were extracted from 34 root-canal-treated teeth with post-treatment apical periodontitis.Each sample was separated into 2 different tubes.PMA was added to one of the tubes,and the other was left untreated.Then,DNA extraction and qPCR were performed.Results:E.faecalis was found in 20 of the 34 samples(58.8%).In PMA treated and none-treaten samples the Ct value of E.faecalis was 25.1 2 ±2.04 and 24.62 ± 2.02 respectively(P =0.001 ).Conclusion:PMA may be feasible in differentiating viable and dead Enterococcus faecalis cells.

Antimicrobial Activity of Tetradenia riparia (Hochst.) Lamiaceae, a Medicinal Plant from Tanzania.

Aims: To evaluate the antibacterial activity of Tetradenia riparia crude extracts against Escherichia coli, Staphylococcus aureus and Enterococcuss faecalis. The phytochemicals that are responsible for the bioactivity were also screened. Study Design: In vitro assay of antibacterial properties. Place and Duration of Study: Samples were collected from Njari village at Uru North in Moshi district located in north eastern Tanzania. Extraction and phytochemical analyses were conducted at the College of Pharmacy and Nutrition, University of Saskatchewan, Saskatoon, Canada. Antimicrobial assay was carried out at Department of Microbiology, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Canada between March 2013 and August 2013. Methodology: Agar well diffusion test was used to determine antimicrobial activity of the plant extracts. Ethanol, methanol, hexane and distilled water were used as extracting solvents. These extracting solvents were removed by vacuo evaporator. The resulting concentrated gummy-like materials were dissolved in Dimethysulfoxide (10% DMSO). Chemical tests were used to determine the group of phytochemicals present in the sample extracts. Results: Sensitivity testing results indicated that S. aureus was found to be more sensitive than E. coli and E. faecalis. Tetradenia riparia methanolic extracts from the root were the most active with zone of inhibition values of 29.33±0.88mm, 21.33±0.33mm and 20.0±1.0mm in diameter against S. aureus, E. faecalis and E. coli respectively. The relative inhibitory zone diameter (RIZD) was calculated. The highest percentage values of relative inhibition zone diameter of 84±5.06% (S. aureus) and 76±6.86% (E. coli) were demonstrated by T. riparia root methanolic extracts. However, T. riparia leaf and root extracts using hexane as well as leaf extracts using water did not show any antibacterial activity against E. faecalis. Root methanolic and ethanolic extracts demonstrated the minimum inhibitory concentrations ranging from 1.25mg/ml to 5.00mg/ml. Phytochemical screening of crude extracts from leaf and root of T. riparia revealed the presence of alkaloids, flavonoids, phenolics, saponins, tannins and sterols. Conclusion: The study findings suggest likelihood of designing and developing potentially active antibacterial drug from T. riparia. Further studies should concentrate on the investigations of not only leaf but also the root part of the plant.

Investigation of virulence factors of Enterococcus faecalis strains isolated in secondary/ persistent infections / Investigação de fatores de virulência de estirpes de Enterococcus faecalis isoladas em infecções secundárias persistentes

More virulent strains may result from the acquisition of genes by genetic exchange, pathogenicity islands in several species encoding toxins, adhesion factors and other factors associated with virulence. The aim of this study was to investigate the prevalence of E. faecalis strains in secondary endodontic/ persistent using endodontic infection by culture and PCR technqiues; and to investigate for the presence of virulence factor genes of gelatinase (gelE), cytolysin activator (Cyla), surface adhesion of Enterococcus (ESP) and collagen adhesin of Enterococcus (ACE). Material and methods: Microbial samples were obtained from 12 teeth with secondary/ persistent endodontic infection showing apical periodontitis. Culture techniques were used including serial dilution, plating, incubation, and biochemical identification. For PCR detection, samples were analyzed using a species-specific primer of the 16S rDNA and the downstream intergenic spacer region. Results: Culture and PCR detected the test species in 3/12 (25%) and 5/12 (41.6%) of teeth,respectively. A total of 38 Enterococcus faecalis strains were isolated and submitted to the virulence factor genes analysis. PCR products consistent with genes encoding surface adhesion (ESP), gelatinase (gelE) and collagen binding antigen (ACE) were found in 26/38 (68%), 31/38 (81%) and 38/38 (100%) of the isolates. The Cytolysin activator (Cyla) gene was not recovered from E. faecalis isolates. Conclusions: In conclusion, the present study revealed by culture and molecular methods revealed a high prevalence of E. faecalis in teeth with secondary/ persistent endodontic infection. Moreover, of a clinical relevance, we found different E. faecalis strains carrying different virulence determinants.

Objetivos: O objetivo deste estudo foi investigar a prevalência de cepas de E. faecalis em canais com infecções endodônticas secundárias/persistentes por meio de cultura e PCR, além de analisar a presença de fatores de virulência genéticos como: gelatinase (gelE), ativador de citolisina (Cyla), adesina de superfície (ESP) e adesina de colágeno (ACE). Material e métodos: Foram coletadas amostras de 12 canais radiculares com infecção endodôntica secundária/persistente e presença de lesão periapical. Para a cultura microbiológica foi realizada diluição em série, incubação e identificação bioquímica dos microrganismos, enquanto que no PCR as amostras foram analisadas através de primers específicos 16S rDNA. Os casos com presença de Enterococcus faecalis foram selecionadas para realização de análise quanto aos fatores de virulência: gelE, Cyla, ESP e ACE. Resultados: Enterococcus faecalis foi detectado através de cultura e PCR em 3/12 (25%) e 5/12 (41,6%) dos casos, respectivamente. No total, foram isoladas 38 amostras com presença de E. faecalis. Os produtos de PCR consistentes com os genes ESP, gelE e ACE foram encontrados em 26 /38 (68%), 31 /38 (81%) e 38/38 (100%) dos isolados. Cyla não foi recuperado a partir de E. faecalis em nenhum dos isolados. Conclusões: O presente estudo revelou alta prevalência de E. faecalis em dentes com infecção endodôntica secundária/ persistente. Estes microrganismos apresentaram elevado índice de diferentes fatores de virulência.

Antibacterial efficacy of sodium hypochlorite with a novel sonic agitation device.

Objective : The aim of this study is to evaluate the antibacterial efficacy of 3% sodium hypochlorite (NaOCl) with sonic agitation devices. Materials and Methods: Fifty extracted human single-rooted teeth were collected. Canals were prepared up to Profile #35/0.06. Forty teeth were inoculated with Enterococcus faecalis and incubated for 24 hours. The teeth were then divided into five groups based on the agitation device used. Group 1- was agitated with EndoActivator . Group 2 was agitated with Waterpik Power Flosser. In Group 3, Waterpik flosser was used with a nickel-titanium file. Group 4 (positive control) was not subjected to any agitation, and Group 5 -was the negative control. Superficial and deep dentinal shavings were collected from coronal and apical halves and sowed in brain heart infusion (BHI) agar to evaluate colony-forming units (CFU/mL). Results: Krusal Wallis, analysis of variance (ANOVA) and Mann-Whitney U test showed that Group 3 showed a smaller number of CFUs/mL Group 2 showed a relatively greater number of CFU/mL. The deep apical dentin areas showed more number of CFUs relative to other sites. Conclusion: The nickel-titanium file attached to the Waterpik Power Flosser system was found to be as effective as the EndoActivator in antibacterial efficacy against E.faecalis.

Efficacy of passive ultrasonic irrigation with natural irrigants (Morinda citrifolia juice, Aloe Vera and Propolis) in comparison with 1% sodium hypochlorite for removal of E. faecalis biofilm: An in vitro study.

Aim: Present study evaluated the efficacy of natural derivative irrigants, Morinda citrifolia juice (MCJ), Aloe Vera and Propolis in comparison to 1% sodium hypochlorite with passive ultrasonic irrigation for removal of the intraradicular E. faecalis biofilms in extracted single rooted human permanent teeth. Materials and Methods: Biofilms of E. faecalis were grown on the prepared root canal walls of 60 standardized root halves which were longitudinally sectioned. These root halves were re-approximated and the samples were divided into five groups of twelve each. The groups were, Group A (1% NaOCl), Group B (MCJ), Group C (Aloe vera), Group D (Propolis) and Group E (Saline). These groups were treated with passive ultrasonic irrigation (PUI) along with the respective irrigants. The root halves were processed for scanning electron microscopy. Three images (X2.5), coronal, middle and apical, were taken for the twelve root halves in each of the five groups. The images were randomized and biofilm coverage assessed independently by three calibrated examiners, using a four-point scoring system. Results: 1% NaOCl with passive ultrasonic irrigation (PUI) was effective in completely removing E. faecalis biofilm and was superior to the natural irrigants like MCJ, Aloe vera and Propolis tested in this study. Conclusion: 1% NaOCl used along with passive ultrasonic irrigation was effective in completely removing E. faecalis biofilm when compared to natural irrigants (MCJ, Aloe Vera and Propolis).

A preliminary study of the antibacterial potential of cetylpyridinium chloride in root canals infected by E. faecalis

The aim of this preliminary study was to verify the antibacterial potential of cetylpyridinium chloride (CPC) in root canals infected by Enterococcus faecalis. Forty human maxillary anterior teeth were prepared and inoculated with E. faecalis for 60 days. The teeth were randomly assigned to the following groups: 1: Root canal preparation (RCP) + 0.1% CPC with positive-pressure irrigation (PPI, Conventional, NaviTip®); 2: RCP + 0.2% CPC PPI; 3: RCP + 2.5% NaOCl PPI; 4: RCP + 2.5% NaOCl with negative-pressure irrigation system (NPI, EndoVac®); 5: Positive control; and 6: Negative control. Four teeth of each experimental group were evaluated by culture and 4 by scanning electron microscopy (SEM). In all teeth, the root canals were dried and filled with 17% EDTA (pH 7.2) for 3 min for smear layer removal. Samples from the infected root canals were collected and immersed in 7 mL of Letheen Broth (LB), followed by incubation at 37°C for 48 h. Bacterial growth was analyzed by turbidity of culture medium and then observed with a UV spectrophotometer. The irrigating solutions were further evaluated for antimicrobial effect by an agar diffusion test.The statistical data were treated by means, standard deviation, Kruskal-Wallis test and analysis of variance. Significance level was set at 5%. The results showed the presence of E. faecalis after root canal sanitization. The number of bacteria decreased after the use of CPC. In the agar diffusion test, CPC induced large microbial inhibition zones, similar to 2% chlorhexidine and large than 2.5% NaOCl. In conclusion, cetylpyridinium chloride showed antibacterial potential in endodontic infection with E. faecalis.

O objetivo deste estudo preliminar foi verificar o potencial antibacteriano de cloreto de cetilpiridínio (CCP) em canais radiculares infectados por E. faecalis. Quarenta dentes anteriores de humanos foram preparados e inoculados com E. faecalis por 60 dias. Os dentes foram aleatoriamente distribuídos como se segue: 1. Preparo do canal radicular (PCR) + CCP 0,1% com sistema de pressão positiva de irrigação (PPI, convencional, Navitip®); 2. PCR + CPC 0,2% PPI; 3. PCR + NaOCl 2,5% PPI, 4. PCR + NaOCl 2,5% com sistema de pressão negativa de irrigação (PNI, EndoVac®); 5 e 6. Controles positivos e negativos. Quatro dentes de cada grupo experimental foram avaliados por cultura e quatro por microscopia eletrônica de varredura (MEV). Em todos os dentes, os canais foram secos e preenchidos com EDTA 17% (pH 7,2) durante 3 min. As amostras dos canais radiculares infectados foram coletadas e imersas em 7 mL Letheen Broth (LB), seguido de incubação a 37° C durante 48 h. O crescimento bacteriano foi analisado pela turvação do meio de cultura, e mensurados por meio de um espectrofotometro (UV). As soluções irrigantes foram ainda avaliadas em teste de difusão em ágar. A análise estatística utilizou de média, desvio padrão,teste de Kruskal-Wallis e análise de variância. O nível de significância foi de 5%. Os resultados mostraram a presença de E. faecalis posterior ao processo de desinfecção do canal radicular. O cloreto de cetilpiridínio mostrou reduzir o número de bactérias. No teste de difusão em ágar, o CPC determinou inibição microbiana, com resultados semelhantes à CHX a 2% e maiores do que o hipoclorito de sódio a 2,5%. O cloreto de cetilpiridínio demonstrou potencial antibacteriano em infecção endodôntica por E. faecalis.

Isolation and characterization of potential lactic acid bacteria (LAB) from freshwater fish processing wastes for application in fermentative utilisation of fish processing waste

Proteolytic and/or lipolytic lactic acid bacteria (LAB) were isolated from visceral wastes of different fresh water fishes. LAB count was found to be highest in case of visceral wastes of Mrigal (5.88 log cfu/g) and lowest in that of tilapia (4.22 log cfu/g). Morphological, biochemical and molecular characterization of the selected LAB isolates were carried out. Two isolates FJ1 (E. faecalis NCIM5367) and LP3 (P. acidilactici NCIM5368) showed both proteolytic and lipolytic properties. All the six native isolates selected for characterization showed antagonistic properties against several human pathogens. All the native isolates were sensitive to antibiotics cephalothin and clindamycin; and, resistant to cotrimoxazole and vancomycin. Considering individually, P. acidilactici FM37, P. acidilactici MW2 and E. faecalis FD3 were sensitive to erythromycin. The two strains FJ1 (E. faecalis NCIM 5367) and LP3 (P. acidilactici NCIM 5368) that had both proteolytic and lipolytic properties have the potential for application in fermentative recovery of lipids and proteins from fish processing wastes.

Isolation and characterization of potential lactic acid bacteria (LAB) from freshwater fish processing wastes for application in fermentative utilisation of fish processing waste

Proteolytic and/or lipolytic lactic acid bacteria (LAB) were isolated from visceral wastes of different fresh water fishes. LAB count was found to be highest in case of visceral wastes of Mrigal (5.88 log cfu/g) and lowest in that of tilapia (4.22 log cfu/g). Morphological, biochemical and molecular characterization of the selected LAB isolates were carried out. Two isolates FJ1 (E. faecalis NCIM5367) and LP3 (P. acidilactici NCIM5368) showed both proteolytic and lipolytic properties. All the six native isolates selected for characterization showed antagonistic properties against several human pathogens. All the native isolates were sensitive to antibiotics cephalothin and clindamycin; and, resistant to cotrimoxazole and vancomycin. Considering individually, P. acidilactici FM37, P. acidilactici MW2 and E. faecalis FD3 were sensitive to erythromycin. The two strains FJ1 (E. faecalis NCIM 5367) and LP3 (P. acidilactici NCIM 5368) that had both proteolytic and lipolytic properties have the potential for application in fermentative recovery of lipids and proteins from fish processing wastes.

Ultrasonic irrigation in the removal of smear layer and Enterococcus faecalis from root canals

Aim: This study evaluated both smear layer removal and reduction of Enterococcus faecalis after instrumentation with ultrasonic irrigation. Methods: Root canals were experimentally inoculated with E. faecalis for 20 days and microbiological samples were collected before and after chemomechanical preparation by using sterilized absorbent paper points. The irrigation solutions used were NaOCl 2.5% and EDTA 17%. In Group 1 (G1), conventional irrigation was used, whereas in Group 2 (G2) ultrasonic irrigation was performed. In group 3 (control), root canals were irrigated with distilled water. The samples were inoculated in BHI broth and turbidity was observed after 48 h to evaluate the reduction in the number of bacteria. Residual smear layer was examined by scanning electron microscopy (SEM). Results: The results showed no significant differences between ultrasonic and conventional irrigation. Conclusions: It was concluded that the level of disinfection and cleanliness of root canals achieved with ultrasonic irrigation is comparable to that obtained by conventional methods.

Characterization of Vancomycin-Resistant Enterococci Isolated from Stools and Their Acquisition of Vancomycin Resistance

We have isolated 6 vancomycin resistant (VR) Enterococcus faecium and 5 VR-E. gallinarum. Vancomycin resistant enterococcus (VRE) isolates were resistant to multi-drugs, but susceptible to linezolid and quinupristin/dalfopristin. VRE isolates showed 10 VanA phenotypes and 1 VanB phenotype (E. gallinarum). However, all of them showed vanA genotype. vanA gene was detected on both genomic and plasmid DNA from all VRE isolates. Almost of VR-E. faecium had IS1216V which is worldwide type and almost of VR-E. gallinarum had IS1542 which is European type. IS1216V and IS1542 genes were not related with antibiotic types of VRE. Copy numbers of vanA were decreased in VRE with IS1216V or IS1542 but not in VRE with both ISs in broth without vancomycin. The copy numbers of vanA were significantly decreased in VanB phenotype of VRE with IS1542 in broth without vancomycin. Copy numbers of vanA were recovered in the presence of vancomycin. Growth time of reference E. faecium is faster than that of reference E. faecalis when cultured in the broth containing vancomycin. Reference strains cultured in the broth containing vancomycin showed intermediate resistance or resistance to antibiotics without acquisition of van genes. Naturally, multidrug-resistant E. faecium might be fast adapted in the presence of vancomycin compared to E. faecalis. Taken together, VanA phenotype E. gallinarum as well as E. feacium have been increasing in nosocomial infection and showed acquired inducible resistance. E. faecium and E. faecalis showed intermediate resistance in long exposure of vancomycin without acquisition of vanA.

Eficácia da clorexidina em infecções endodônticas: revisão sistemática / Efficacy of chlorhexidine in endodontic infections: systematic review

Avaliou-se em estudos longitudinais a eficácia da clorexidina em infecções endodônticas detectadas por cultura ou reação em cadeia da polimerase (PCR), por meio de revisão sistemática. As estratégias de busca incluíram buscas eletrônicas (Medline, Embase, Central) e manuais, utilizando-se várias palavras-chave: chlorhexidineand (endodontic* OR endodontic* infection* OR root canal infection*). A busca apresentou 196 artigos, sendo que dos 48 estudos in vivo, sete satisfizeram os critérios de inclusão. A combinação de resultados com vistas à estruturação de uma meta-análise não foi possível em função da heterogeneidade dos estudos. O emprego da clorexidina como irrigante durante o preparo de canais radiculares infectados mostrou reduzir a microbiota endodôntica remanescente.

Influence of the length of remaining root canal filling and post space preparation on the coronal leakage of Enterococcus faecalis / Influência do comprimento do material obturador remanescente no canal radicular e preparo do espaço protético na infiltração coronária do Enterococcus faecalis

This study evaluated the sealing ability of different lengths of remaining root canal filling and post space preparation against coronal leakage of Enterococcus faecalis. Forty-one roots of maxillary incisors were biomechanically prepared, maintaining standardized canal diameter at the middle and coronal thirds. The roots were autoclaved and all subsequent steps were undertaken in a laminar flow chamber. The canals of 33 roots were obturated with AH Plus sealer and gutta-percha. The root canal fillings were reduced to 3 predetermined lengths (n=11): G1=6 mm, G2=4 mm and G3=2 mm. The remaining roots served as positive and negative controls. Bacterial leakage test apparatuses were fabricated with the roots attached to Eppendorf tubes keeping 2 mm of apex submerged in BHI in glass flasks. The specimens received an E. faecalis inoculum of 1 x 107 cfu/mL every 3 days and were observed for bacterial leakage daily during 60 days. Data were submitted to ANOVA, Tukey's test and Fisher's test. At 60 days, G1 (6 mm) and G2 (4 mm) presented statistically similar results (p>0.05) (54.4% of specimens with bacterial leakage) and both groups differed significantly (p<0.01) from G3 (2 mm), which presented 100% of specimens with E. faecalis leakage. It may be concluded that the shortest endodontic obturation remnant leaked considerably more than the other lengths, although none of the tested conditions avoids coronal leakage of E. faecalis.

Este estudo avaliou a capacidade de diferentes de remanescentes de material obturador do canal radicular e preparo do espaço protético na infiltração do Enterococcus faecalis. Quarenta e uma raízes de incisivos superiores foram preparadas biomecanicamente, mantendo-se diâmetro padronizado nos terços médio e cervical. As raízes foram autoclavadas e todos os passos subseqüentes foram realizados em capela de fluxo laminar. Os canais de 33 raízes foram obturadors com AH Plus e guta-percha. As obturações foram reduzidas a 3 comprimentos (n=11): G1=6 mm, G2=4 mm e G3=2 mm. As raízes remanescentes serviram de controles positivo e negativo. O dispositivo para testar a microinfiltração bacteriana foi confeccionado com as raízes fixas a Eppendorfs, mantendo-se 2 mm do ápice submergido em vidro contendo BHI. Os dentes receberam o inóculo de 1 x 10(7) UFC/ml de E. faecalis a cada 3 dias, com observação diária por 60 dias. Os dados obtidos foram submetidos à análise de variância, teste de Tukey e Fisher. Foi possível observar que aos 60 dias, o G1 (6 mm) e G2 (4 mm) apresentou resultados estatisticamente semelhantes (p>0,05) (54% dos espécimes com infiltração bacteriana) e ambos os grupos foram diferentes estatisticamente (p<0,01) do G3 (2 mm), o qual apresentou 100% de espécimes com microinfiltração. Concluiu-se que apesar da infiltração do E. faecalis ter ocorrido em todas as condições testadas, aparentemente houve uma correlação positiva entre o comprimento do remanescente radicular e a eficácia do selamento, uma vez que, a menor a obturação remanescente infiltrou consideravelmente mais que os outros comprimentos.

Evaluation of Enterococcus faecalis removal efficacy of the EndoVac(R) and EndoActivator(R) intracanal irrigation methods / 대한치과보존학회지

The aim of this study was to evaluate endodontic irrigation methods with EndoVac(R) and EndoActivator(R) in the elimination of Enterococcus faecalis from the root canals. Extracted 70 human single-rooted teeth were used. The canals were instrumented by a crown-down technique with .04 taper ProFile to ISO size 40. After the teeth were autoclaved, the canals were inoculated with E. faecalis and incubated for 48 h. The teeth were randomly divided into three experimental groups of 20 teeth each according to canal irrigation methods and two control groups as follows: group 1 - EndoVac(R) group 2 - EndoActivator(R) ; group 3 - Conventional needle irrigation method. After canal irrigation using 2.5% NaOCl, first samples (S1) were taken using sterile paper point. And the canals were filled with sterile brain heart infusion (BHI) broth and incubated for 24 h, then second samples (S2) were taken. The samples were cultured on BHI agar plate to determine the numbers of colony forming units (CFU). In first sampling (S1), only one canal of conventional method among the all experimental groups was positive cultured. In second sampling (S2), EndoVac(R) group showed the least positive culture numbers of E. faecalis. There was statistically significant difference between the EndoVac(R) and conventional needle irrigation methods in the mean value of Log CFU. According to the results of this study, EndoVac(R) showed better efficacy than conventional needle irrigation method in the elimination of E. faecalis from the root canal.