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The aim of this study was to investigate the antimicrobial activity of ethanolic extract and fractions The aim of this study was to investigate the antimicrobial activity of ethanolic extract and the fractions
of Brazilian green propolis (BGP) collected by bees from Baccharis dracunculifolia against 16 oral of Brazilian green of propolis (BGP) collected by Bees from Baccharis dracunculifolia against 16 oral
pathogenic microorganisms. pathogenic microorganisms. BGP was examinated by Reversed-Phase High-Performance Liquid BGP was examinated by Reversed-Phase High-Performance Liquid
Chromatography (RPHPLC) and its absorption spectra was assessed using UV-Spectrophotometer. Chromatography (RPHPLC), and its absorption Spectra was assessed using UV-Spectrophotometer.
Identification of flavonoids and other chemical constituents were carried out using authentic Identification of flavonoids and other Chemical constituents were carried out using authentic
standards. Antimicrobial activity was evaluated by agar diffusion and dilution method. Antimicrobial activity was evaluated by diffusion and dilution that method. The results The results
indicate that all microorganisms tested were susceptible to BGP. indicate that all microorganisms Tested were susceptible to BGP. None of the essayed fractions None of the essayed fractions
(Coumaric acid, Kaempferol, Pinobanskin-3-acetate, Chrysin, Galangin, Kaempferide, and Artepillin (Coumaric acid, Kaempferol, Pinobanskin-3-acetate, Chrysin, Galangin, Kaempferide, and Artepillin
C) was more active than the extract, suggesting a synergistic effect of propolis constituents for the C) was more active than the extract, suggesting a synergistic effect of the constituents of propolis
antimicrobial activity. antimicrobial activity.
Propolis is a natural resinous hive product used by the bees to protect the hive against the invasion of Propolis is a resinous hive natural products used by the Bees in the hive to protect against the invasion of
microorganisms and is considered to be a natural antibiotic1. microorganisms, and is considered to be a natural antibiotic1. It has also been used extensively in folk It has also been extensively used in folk
medicine by the Brazilian population for several years. medicine by the Brazilian population for several years. Many beekeepers, pharmacists and local Many beekeepers, pharmacists and local
laboratories in Brazil produce a great variety of propolis derivatives for medicinal use. Laboratories in Brazil produce a great Variety of propolis of derivatives for medicinal use. Beekeepers
commonly chew raw propolis to treat mouth and upper digestive track infections. Chew raw comments of propolis to treat mouth and upper digestive track infections. Available literature Available literature
indicates that few antimicrobial studies have been carried out using Brazilian propolis and there are indicates that antimicrobial few studies have been carried out using Brazilian of propolis, and there are
only a few reports documenting the chemical constituents and their biological activities2,3. only a few reports documenting the Chemical constituents and their Biological activities2, 3. The aim of The aim of
this study was to evaluate the antibacterial and antifungal activity of Brazilian green propolis extract this study was to evaluate the antibacterial and antifungal activity of Brazilian green extract of propolis
and fractions against Candida spp., Gram positive and Gram negative oral pathogenic bacteria. and the fractions against Candida spp., Gram positive and Gram negative oral pathogenic bacteria.

Crude Brazilian green propolis was obtained from an apicultural region of Minas Gerais State, Brazil Crude Brazilian Green of propolis was obtained from an apicultural region of Minas Gerais State, Brazil
by PharmaNectar®(REF. LOT. SBN97). by PharmaNectar ® (REF. LOT. SBN97). 100g of crude propolis was kept in a freezer for 24 h and 100g of Crude of propolis was kept in a freezer for 24 h and
powdered in a blender. powdered in a blender. After dissolving, ethanolic extracts of Brazilian green propolis (BGP) was After dissolving, ethanolic extracts of propolis of Brazilian green (BGP) was
filtered through Sigma nº1 filter paper. filtered through Sigma n º 1 filter paper. The filtered extract was concentrated under vacuum to furnish The extract was filtered concentrate under vacuum to Furnish
62g of a crude extract. 62g of a Crude extract. Analysis of flavonoids from ethanolic extracts of bud and unexpanded leaf Analysis of flavonoids from ethanolic extracts of Bud and Leaf unexpanded
exudates and ethanolic extracts of propolis was performed by RPHPLC2 with a chromatograph exudates and ethanolic extracts of propolis of RPHPLC2 was performed by a chromatograph
equipped with a YMC PACK ODS-A column (RP-18, column size 4.6 x 250 mm; particle size 5μm) equipped with a YMC PACK-ods A column (RP-18, column size 4.6 x 250 mm; particle size 5μm)
and Photodiode Array Detector (SPD- M10A, Shimazu Co., Japan). and Photodiode Array Detector (SPD-M10A, Shimazu Co.., Japan). The column was eluted by using The column was eluted by using
a linear gradient of water (solvent A) and methanol (solvent B) starting with 30% B (0-15 min), and a linear gradient of water (solvent A) and methanol (solvent B) starting with 30% B (0-15 min), and
increasing to 90% B (15-75min), held at 90% B (75-95 min) and decreasing to 30% B (95-105 min) Increasing to 90% B (15-75min), held at 90% B (75-95 min) and decreasing to 30% B (95-105 min)
with a solvent flow rate of 1 mL/min and detection with a diode array detector. solvent with a flow rate of 1 mL / min and detection with a diode array detector. Chromatograms were Chromatograms were
recorded at 268nm. recorded at 268nm. The ethanolic extract of propolis was measured the absorption spectra, using UVSpectrophotometer. The ethanolic extract of propolis of absorption was measured in the Spectra, using UVSpectrophotometer.
Identification of flavonoids and other chemical constituents were carried out using authentic Identification of flavonoids and other Chemical constituents were carried out using authentic
standards purchased from Extrasythese Co. standards purchased from Extrasythese Co.. (France). The authentic standard of 3,5-dipremyl-4- The authentic standard of 3.5-dipremyl-4 -
hydroxycinnamic acid (artepillin C) was donated from Hayashibara Biochemical Laboratory hydroxycinnamic acid (artepillin C) was donated from Hayashibara Biochemical Laboratory
(Okayama, Japan). (Okayama, Japan). The authentic standard of Pinobanksin and Pinobanksin-3-acetate were donated The authentic standard of Pinobanksin and Pinobanksin-3-acetate were donated
from Dr. from Dr. E. Wollenweber (Institue für Botanik, Technishe Hochschule Darmstadt, Germany). Wollenweber (Institue für Botanik, Technishe Hochschule Darmstadt, Germany). The
minimal inhibitory concentration (MIC) was defined as the lowest concentration of propolis in which minimum inhibitory Concentration (MIC) was defined as the Lowest of the Concentration of propolis in which
no bacterial growth was detected. no bacterial growth was Detected. Determination of MICs by the agar dilution method was Determination of MICs by the dilution method was to
performed, following the serial concentrations of BGP and different fractions were achieved (%v/v) performed, following the serial concentrations of BGP and the different fractions were achieved (% v / v)
in plates containing Brucella agar (Oxoid), as follows: 0.1, 0.2, 0.4, 0.8, 1.75, 3.5, 7.0 and 14.0. in the plates containing Brucella to (Oxoid), as follows: 0.1, 0.2, 0.4, 0.8, 1.75, 3.5, 7.0 and 14.0. Each
antimicrobial test also included plates containing the culture medium plus ethanol, in order to obtain antimicrobial test also included plates containing the culture medium plus ethanol, in order to obtain
a control of the solvent antibacterial effect4,5. a solvent control of the antibacterial effect4, 5. The antimicrobial and antifungal susceptibility test for The antimicrobial and antifungal susceptibility test for
Streptococcus mutans (ATCC 70069), Strepetococcus sanguis (ATCC 10557), Lactobacillus casei Streptococcus mutans (ATCC 70069), Strepetococcus sanguis (ATCC 10557), Lactobacillus casei
(ATCC 393), Tanerella forsythensis (ATCC 700191), Bacteroides fragilis (ATCC 25285), (ATCC 393), Tanerella forsythensis (ATCC 700191), Bacteroides fragilis (ATCC 25285),
Staphylococcus aureus (ATCC 12692), Fusobacterium necrophorum (ATCC 25286), Actinobacillus Staphylococcus aureus (ATCC 12692), Fusobacterium necrophorum (ATCC 25286), Actinobacillus
actinomycetemcomitans (ATCC 33384), Porphyromonas gingivalis (ATCC 33277), Fusobacterium actinomycetemcomitans (ATCC 33384), Porphyromonas gingivalis (ATCC 33277), Fusobacterium
nucleatum (ATCC 23726), C. nucleatum (ATCC 23726), C. albicans (ATCC 18804), C. albicans (ATCC 18804), C. tropicalis (ATCC 750), C. tropicalis (ATCC 750), C. glabrata
(ATCC 2001), C. (ATCC 2001), C. parapsilosis (ATCC 22019), C. parapsilosis (ATCC 22019), C. krusei (ATCC 2340) and C. krusei (ATCC 2340) and C. guilliermondii (ATCC guilliermondii (ATCC
201935) were studied with reference microdilution method following the NCCLS M27-AZ Standards 201935) were studied with reference microdilution method, following the NCCLS M27-AZ Standards
by using RPMI 1640 medium (Sigma-USA) with L-glutamine and phenol red and without sodium by using RPMI 1640 medium (Sigma-USA) with L-glutamine, and phenols, red and without sodium
bicarbonate6, 7. bicarbonate6, 7. Yeast suspension were inoculated into microplate wells which contained 1/64-1/8000 Yeast suspension were inoculated into microplate Wells which contained 1/64-1/8000
dilution of BGP and fractions solutions. dilution of BGP and fractions solutions. Microplates were evaluated after incubation at 37ºC for 48 h. Microplates were evaluated after incubation at 37 º C for 48 h.
Sterile blank disks (CECON - São Paulo - Brazil) were soaked in 20 μl of the BGP solution, 20μl of Sterile blank disks (CECON - São Paulo - Brazil) were soaked in 20 μl of the BGP solution, 20μl of
each component Coumaric acid, kaempferol, Pinobanskin-3-acetate, chrysin, galangin, kaempferide, each component Coumaric acid, kaempferol, Pinobanskin-3-acetate, chrysin, galangin, kaempferide,
and artepillin C, and applied to the agar surface previously seeded with the microorganism. artepillin and C, and Applied to the surface that previously seeded with the microorganism.
Positive and negative controls of the discs containing 30μg of tetracycline, Nystatin 30mg, and 20μl Positive and negative controls of the discs containing 30μg of tetracycline, Nystatin 30mg, and 20μl
of Ethanol 93,2ºC were used. Ethanol of 93.2 º C were used. After 48 hours of incubation at 37ºC, the diameters of the inhibition After 48 hours of incubation at 37 º C, the diameters of the inhibition
zones were measured and compared. Zones were measured and the company. The results of the diameters of the inhibition zones were The results of the diameters of inhibition were Zones
reported as Means + Standard Deviation (M±SD). reported as Means + Standard Deviation (M ± SD). The inhibitory ability of the various propolis The inhibitory Yusuf of the download of propolis
solutions tested on the oral pathogenic bacteria and fungi was compared with nonparametric Kruskal- Tested solutions on the oral pathogenic bacteria, and it was a company function with nonparametric Kruskal -
Wallis test. Wallis test. Differences of the level p<0.05 were considered to be significant. Differences of the level of p <0:05 were considered to be significant.