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      Antibacterial activity of a plant extract and its potential for disinfecting gutta-percha cones

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          Abstract

          This study evaluated the antibacterial activity of Rosmarinus officinalis extract and its potential for disinfecting guttapercha (GP) cones. In the first experiment, a hydro-alcoholic extract of Rosmarinus officinalis (leaves) in a dilution ratio of 10% m/v was tested against Enterococcus faecalis by using the disk diffusion method. Positive and negative controls were 70% cereal alcohol and antibiotics, respectively. The procedures were performed in triplicate, and the diameters of the zones of growth inhibition were measured with a caliper after 24 h at 37ºC. In the second experiment, the disinfection procedures were evaluated on GP cones artificially contaminated with Enterococcus faecalis. The R. officinalis extract was compared with 2% chlorhexidine digluconate and 2.5% sodium hypochlorite, using a direct exposure test (5 min treatment). Sterilized and non-disinfected cones were used as negative and positive controls, respectively. After 24 h of incubation, bacterial counts were taken. For both experiments, the data were statistically analyzed by Kruskall-Wallis and Tukey's tests (p<0.05). The plant extract produced zones of inhibition comparable to those of tested antibiotics. Significant GP cone disinfection was verified with all disinfectant solutions, with no significant difference between them. R. officinalis extract showed bactericidal effect on Enterococcus faecalis and capacity to disinfect GP cones contamined with it.

          Translated abstract

          Este estudo avaliou a atividade antibacteriana do extrato de Rosmarinus officinalis e do potencial deste extrato vegetal para a desinfecção de cones de guta-percha (GP). No primeiro experimento, o extrato hidroalcoólico de Rosmarinus officinalis (folhas), em diluição de 10% m/v, foi testado contra Enterococcus faecalis, utilizando o método de discodifusão em ágar. Os controles positivos e negativos foram o álcool de cereais 70% e antibióticos, respectivamente. Os procedimentos foram realizados em triplicata e os diâmetros de halos de inibição foram mensurados com um paquímetro, após 24 horas a 37 º C. No segundo experimento, os procedimentos de desinfecção foram avaliados em cones de GP artificialmente contaminados com Enterococcus faecalis. O extrato de R. officinalis foi comparado com o gluconato de clorexidina a 2% e com hipoclorito de sódio a 2,5% através de um teste de exposição direta por 5 minutos. Cones esterili - zados e cones não desinfectados foram utilizados como controles negativo e positivo, respectivamente. Após 24h de incubação, as contagens bacterianas foram enumeradas. Em ambos os experimentos, os dados foram analisados estatisticamente pelos testes de Kruskall-Wallis e Tukey (p <0,05). O extrato vegetal apresentou halos de inibição semelhantes aos antibióticos testados. Expessiva descontaminação dos cones foi verificada com todas as soluções desinfetantes, sem dife - renças significativas entre elas. O extrato de R. officinalis mostrou efeito bactericida sobre Enterococcus faecalis e capacidade de desinfetar cones de GP contaminados com este microrganismo.

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          Evaluation of diffusion and dilution methods to determine the antibacterial activity of plant extracts.

          The aim of this study was to evaluate diffusion and dilution methods for determining the antibacterial activity of plant extracts and their mixtures. Several methods for measurement of the minimal inhibitory concentration (MIC) of a plant extract are available, but there is no standard procedure as there is for antibiotics. We tested different plant extracts, their mixtures and phenolic acids on selected gram-positive (Staphylococcus aureus, Bacillus cereus, and Listeria monocytogenes) and gram-negative bacteria (Escherichia coli O157:H7, Salmonella Infantis, Campylobacter jejuni, Campylobacter coli) with the disk diffusion, agar dilution, broth microdilution and macrodilution methods. The disk diffusion method was appropriate only as a preliminary screening test prior to quantitative MIC determination with dilution methods. A comparison of the results for MIC obtained by agar dilution and broth microdilution was possible only for gram-positive bacteria, and indicated the latter as the most accurate way of assessing the antimicrobial effect. The microdilution method with TTC (2,3,5-triphenyl tetrazolium chloride) or INT (2-p-iodophenyl-3-p-nitrophenyl-5-phenyl tetrazolium chloride) to indicate the viability of aerobic bacteria was found to be the best alternative approach, while only ATP determination was appropriate for microaerophilic Campylobacter spp. Using survival curves the kinetics of bacterial inactivation on plant extract exposure was followed for 24h and in this way the MIC values determined by the microdilution method were confirmed as the concentrations of extracts that inhibited bacterial growth. We suggest evaluation of the antibacterial activity of plant extracts using the broth microdilution method as a fast screening method for MIC determination and the macrodilution method at selected MIC values to confirm bacterial inactivation. Campylobacter spp. showed a similar sensitivity to plant extracts as the tested gram-positive bacteria, but S. Infantis and E. coli O157:H7 were more resistant. Copyright 2010 Elsevier B.V. All rights reserved.
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            Composition and antimicrobial activity of essential oils from aromatic plants used in Brazil

            Essential oils from aerial parts of Mentha piperita, M. spicata, Thymus vulgaris, Origanum vulgare, O. applii, Aloysia triphylla, Ocimum gratissimum, O. basilicum were obtained by steam destillation using a Clevenger-type system. These oils were screened for antibacterial and anti-Candida albicans activity using bioautographic method. Subsequently, minimal inhibitory concentration from oils was determined by microdilution method. Most essential oil studied were effective against Enterococcus faecium and Salmonella cholerasuis. Aloysia triphylla and O. basilicum presented moderate inhibition against Staphylococcus aureus while only A. tryphila and M. piperita were able to control the yeast Candida albicans. The oils were analyzed by GC and GC-MS techniques in order to determine the majoritary compounds.
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              Antimicrobial activity of Rosmarinus officinalis against oral pathogens: relevance of carnosic acid and carnosol.

              The in vitro inhibitory activity of crude EtOH/H2O extracts from the leaves and stems of Rosmarinus officinalis L. was evaluated against the following microorganisms responsible for initiating dental caries: Streptococcus mutans, S. salivarius, S. sobrinus, S. mitis, S. sanguinis, and Enterococcus faecalis. Minimum inhibitory concentrations (MIC) were determined with the broth microdilution method. The bioassay-guided fractionation of the leaf extract, which displayed the higher antibacterial activity than the stem extract, led to the identification of carnosic acid (2) and carnosol (3) as the major compounds in the fraction displaying the highest activity, as identified by HPLC analysis. Rosmarinic acid (1), detected in another fraction, did not display any activity against the selected microorganisms. HPLC Analysis revealed the presence of low amounts of ursolic acid (4) and oleanolic acid (5) in the obtained fractions. The results suggest that the antimicrobial activity of the extract from the leaves of R. officinalis may be ascribed mainly to the action of 2 and 3.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                aol
                Acta Odontológica Latinoamericana
                Acta odontol. latinoam.
                Sociedad Argentina de Investigación Odontológica (Buenos Aires, Ciudad Autonoma de Buenos Aires, Argentina )
                1852-4834
                April 2012
                : 25
                : 1
                : 9-13
                Affiliations
                [02] Montes Claros MG orgnameState University of Montes Claros orgdiv1Department of Biology Brazil
                [01] Montes Claros MG orgnameState University of Montes Claros orgdiv1Department of Dentistry Brazil
                [03] Aracaju SE orgnameFederal University of Sergipe orgdiv1Department of Dentistry Brazil
                Article
                S1852-48342012000100002
                90b7b00f-0ffe-4895-96fb-0f01804d4077

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

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                Figures: 0, Tables: 0, Equations: 0, References: 23, Pages: 5
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                SciELO Argentina


                Plant extracts,Enterococcus faecalis,Gutta-percha,Extratos vegetais,Guta-percha

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