In vitro antimicrobial activity of propolis samples from different geographical origins against certain oral pathogens

Propolis is an agent having antimicrobial properties, however, its composition can vary depending on the area where it is collected. In the present study, the antimicrobial activity of five propolis samples, collected from four different regions in Turkey and from Brazil, against nine anaerobic strains was evaluated. Ethanol extracts of propolis (EEP) were prepared from propolis samples and we determined minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of EEP on the growth of test microorganisms by using agar dilution method. All strains were susceptible and MIC values ranged from 4 to 512 microg/ml for propolis activity. Propolis from Kazan-Ankara showed most effective MIC values to the studied microorganisms. MBC values of Kazan-Ankara EEP samples were ranged from 8 to 512 microg/ml. Death was observed within 4 h of incubation for Peptostreptococcus anaerobius and micros and Lactobacillus acidophilus and Actinomyces naeslundii, while 8 h for Prevotella oralis and Prevotella melaninogenica and Porphyromonas gingivalis, 12 h for Fusobacterium nucleatum, 16 h for Veillonella parvula. It was shown that propolis samples were more effective against Gram positive anaerobic bacteria than Gram negative ones. The organic chemical compositions of EEPs were determined by high-resolution gas chromatography coupled to mass spectrometry (GC-MS). The main compounds of EEPs were flavonoids such as pinobanksin, quercetin, naringenin, galangine, chrysin and aromatic acids such as cafeic acid. Because of increased antimicrobial resistance, propolis may be kept in mind in the treatment of oral cavity diseases.     

Antimicrobial effect of Korean propolis against the mutans streptococci isolated from Korean

The aim of this study was to determine the optimal concentration of Korean propolis against clinical isolates of mutans streptococci (MS) from Koreans. The antimicrobial activity was evaluated using the minimum inhibitory concentration (MIC) and time-kill curves against mutans streptococci. The MIC(90) values of propolis for MS were 35 μg/ml. Propolis had a bacteriostatic effect on Streptococcus mutans ATCC 25175(T) and bactericidal effects on Streptococcus sobrinus ATCC 33478(T) at > 2 × MIC (70 μg/ml). These results suggest that the propolis can be used in the development of oral hygiene products for the prevention of dental caries.     

Egyptian propolis: 1-antimicrobial activity and chemical composition of Upper Egypt propolis

The antimicrobial activity of four propolis samples collected from Upper Egypt against Staphylococcus aureus; Escherichia coli and Candida albicans was evaluated. There was a variation in the antimicrobial activity according to the propolis origin. Banisweif propolis showed the highest antimicrobial activity against Staphylococcus aureus, Escherichia coli and Candida albicans, but Fayoum propolis had moderate activity against all tested pathogens. Propolis collected from Assiut and Souhag gave lower antimicrobial activity. Propolis samples were investigated by GC/MS, 71 compounds were identified, 14 being new for propolis. Banisweif propolis is characterized by the presence of 7 caffeate esters and 4 triterpenoids. Fayoum propolis showed the highest amount of lactic acid and the presence of 3 chalcones. But Assiut propolis is characterized by the presence of 4 prenylated coumarates. Souhag propolis is characterized by the presence of 5 aliphatic dicarboxylic acids and some other new compounds to propolis.     

The antimicrobial effects of propolis collected in different regions in the Basque Country (Northern Spain)

The antimicrobial activity of 19 propolis extracts prepared in different solvents (ethanol and propylene glycol) (EEP/PEP), was evaluated against some bacterial and fungal isolates using the agar-well diffusion method. It was verified that all the samples tested showed antimicrobial activity, although results varied considerably between samples. Results revealed that both types of propolis extracts showed highly sensitive antimicrobial action against Gram-positive bacteria and fungi at a concentration of 20% (Staphylococcus aureus, Streptococcus mutans, Candida albicans and Saccharomyces cerevisae) with a minimal inhibitory concentration (MIC) ranging from 0.5 to 1.5 mg/ml, with a moderate effect against Streptococcus pyogenes (MIC from 17 to 26 mg/ml). To our knowledge, this is the first study showing elevated antimicrobial activity against Gram-negative bacteria [Salmonella enterica (MIC from 0.6 to 1.4 mg/ml)] and lesser activity against Helicobacter pylori (MIC from 6 to 14 mg/ml), while Escherichia coli was resistant. This concluded that the Basque propolis had a strong and dose-dependent activity against most of the microbial strains tested, while database comparison revealed that phenolic substances were responsible for this inhibition, regardless of their geographical origin and the solvent employed for extraction. Statistical analysis showed no significant differences (P ≤ 0.05) between EEP and PEP extracts.     

Anticariogenic activity of macelignan isolated from Myristica fragrans (nutmeg) against Streptococcus mutans

The occurrence of dental caries is mainly associated with oral pathogens, especially cariogenic Streptococcus mutans. Preliminary antibacterial screening revealed that the extract of Myristica fragrans, widely cultivated for the spice and flavor of foods, possessed strong inhibitory activity against S. mutans. The anticariogenic compound was successfully isolated from the methanol extract of M. fragrans by repeated silica gel chromatography, and its structure was identified as macelignan by instrumental analysis using 1D-NMR, 2D-NMR and EI-MS. The minimum inhibitory concentration (MIC) of macelignan against S. mutans was 3.9 microg/ml, which was much lower than those of other natural anticariogenic agents such as 15.6 microg/ml of sanguinarine, 250 microg/ml of eucalyptol, 500 microg/ml of menthol and thymol, and 1000 microg/ml of methyl salicylate. Macelignan also possessed preferential activity against other oral microorganisms such as Streptococcus sobrinus, Streptococcus salivarius, Streptococcus sanguis, Lactobacillus acidophilus and Lactobacillus casei in the MIC range of 2-31.3 microg/ml. In particular, the bactericidal test showed that macelignan, at a concentration of 20 microg/ml, completely inactivated S. mutans in 1 min. The specific activity and fast-effectiveness of macelignan against oral bacteria strongly suggest that it could be employed as a natural antibacterial agent in functional foods or oral care products.     

Effect of a New Variety of Apis mellifera Propolis onMutans Streptococci

The effects of a new variety of propolis, from Northeastern Brazil (BA), on growth of mutans streptococci, cell adherence, and water-insoluble glucan (WIG) synthesis were evaluated. Propolis from Southeastern (MG) and Southern (RS) Brazil were also tested as an extension of our previous work. Ethanolic extracts of propolis (EEP) were prepared and analyzed by reversed-phase HPLC. For the antibacterial activity assays, minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of EEPs against Streptococcus mutans, S. sobrinus, and S. cricetus were determined. Cell adherence of S. mutans and S. sobrinus to a glass surface was measured spectrophotometrically at 550 nm. WIG synthesized from sucrose by glucosyltransferase (Gtf) was extracted and quantified by the phenol-sulfuric method. The HPLC profile of the new variety of propolis was entirely different from Southeastern and Southern propolis. Neither flavonoid aglycones nor p-coumaric acid were detected in EEP BA. All EEPs demonstrated biological activities against mutans streptococci; EEP BA showed the highest potency in all in vitro parameters evaluated in this study. The ranges of MIC values were 50 (EEP BA)–400 μg/ml (MG), for S. mutans; and 25 (BA)–400 μg/ml (MG), for S. sobrinus and S. cricetus. The bactericidal concentration of EEPs was four to eight times the MIC values. The adherence of S. mutans and S. sobrinus cells and WIG synthesis were markedly inhibited by EEPs, demonstrating significant inhibition at all concentrations compared with the control (80% ethanol) (p < 0.05). EEP BA showed 80% inhibition of cell adherence and WIG synthesis at concentrations as low as 12.5 and 7.8 μg/ml, respectively. The results show that the new variety of propolis was exceptionally effective in all in vitro parameters tested against mutans streptococci; biological effects of propolis are likely not to be due solely to flavonoids and (hydroxy)cinnamic acid derivatives. Received: 14 February 2000 / Accepted: 8 May 2000     

In vitro antimicrobial activity of propolis and synergism between propolis and antimicrobial drugs

The aim of this study was to investigate antimicrobial properties of ethanolic extract of 13 propolis (EEP) samples from different regions of Serbia against 39 microorganisms (14 resistant or multiresistant to antibiotics), and to determine synergistic activity between antimicrobials and propolis. Antimicrobial activity of propolis samples was evaluated by agar diffusion and agar dilution method. The synergistic action of propolis with antimicrobial drugs was assayed by the disc diffusion method on agar containing subinhibitory concentrations of propolis. Obtained results indicate that EEP, irrespectively of microbial resistance to antibiotics, showed significant antimicrobial activities against Gram-positive bacteria (MIC 0.078%–1.25% of EEP) and yeasts (0.16%–1.25%), while Gram-negative bacteria were less susceptible (1.25&%ndash;>5%). Enterococcus faecalis was the most resistant Gram-positive bacterium, Salmonella spp. the most resistant Gram-negative bacteria, and Candida albicans the most resistant yeast. EEP showed synergism with selected antibiotics, and displayed ability to enhance the activities of antifungals. The shown antimicrobial potential of propolis alone or in combination with certain antibiotics and antifungals is of potential medical interest.     

Identification of a bioactive compound isolated from Brazilian propolis type 6

A prenylated benzophenone, hyperibone A, was isolated from the hexane fraction of Brazilian propolis type 6. Its structure was determined by spectral analysis including 2D NMR. This compound exhibited cytotoxic activity against HeLa tumor cells (IC₅₀ = 0.1756 μM), strong antimicrobial activity (MIC range—0.73–6.6 μg/mL; MBC range—2.92–106 μg/mL) against Streptococcus mutans, Streptococcus sobrinus, Streptococcus oralis, Staphylococcus aureus, and Actinomyces naeslundii, and the results of its cytotoxic and antimicrobial activities were considered good.     

The antimicrobial activity of essential oil from Dracocephalum foetidum against pathogenic microorganisms

A number of essential oils from Mongolian aromatic plants are claimed to have antimicrobial activities. The essential oil of Dracocephalum foetidum, a popular essential oil used in Mongolian traditional medicine, was examined for its antimicrobial activity. Eight human pathogenic microorganisms including B. subtilis, S. aureus, M. lutens, E. hirae, S. mutans, E. coli, C. albicans, and S. cerevisiae were examined. The essential oil of Dracocephalum foetidum exhibited strong antimicrobial activity against most of the pathogenic bacteria and yeast strains that were tested; by both the agar diffusion method and the minimum inhibitory concentration (MIC) assay (MIC range was 26-2592 microg/ml). Interestingly, Dracocephalum foetidum even showed antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA) strains. We also analyzed the chemical composition of the oil by GC-MS and identified several major components, including n-Mentha-1,8-dien-10-al, limonene, geranial, and neral.     

Antimicrobial activity of kaurane diterpenes against oral pathogens

Two kaurane diterpenes, ent-kaur-16(17)-en-19-oic acid (KA) and 15-beta-isovaleryloxy-ent-kaur-16(17)-en-19-oic acid (KA-Ival), isolated from Aspilia foliacea, and the methyl ester derivative of KA (KA-Me) were evaluated against oral pathogens. KA was the most active compound, with MIC values of 10 microg mL(-1) against the following microorganisms: Streptococcus sobrinus, Streptococcus mutans, Streptococcus mitis, Streptococcus sanguinis, and Lactobacillus casei. However, KA did not show significant activity against Streptococcus salivarius and Enterococcus faecalis, with MIC values equal to 100 and 200 microg mL(-1), respectively. Our results show that KA has potential to be used as a prototype for the discovery of new effective anti-infection agents against microorganisms responsible for caries and periodontal diseases. Moreover, these results allow to conclude that minor structural differences among these diterpenes significantly influence their antimicrobial activity, bringing new perspectives to studies on the structure-activity relationship of this type of metabolites with respect to caries and periodontal diseases.     

Egyptian propolis: 2. Chemical composition,antiviral and antimicrobial activities of East Nile Delta propolis

Three propolis samples from East Nile Delta, Egypt were collected. Propolis samples were investigated by GC/MS,103 compounds were identified, 20 being new for propolis. Dakahlia propolis was a typical poplar propolis but it contained two new caffeate esters and two new triterpenoids. Ismailia propolis was characterized by the presence of new triterpenic acid methyl esters and it did not contain any aromatic acids, esters and flavonoids. Sharkia propolis was characterized by the presence of caffeate esters only, some di- and triterpenoids. The antiviral (Infectious Bursal Disease Virus and Reo-Virus) and antimicrobial (Staphylococcus aureus; Escherichia coli and Candida albicans) activities of propolis samples were investigated. Dakahlia propolis showed the highest antiviral activity against Infectious Bursal Disease Virus (IBDV) and the highest antibacterial activity against Escherichia coli and the highest antifungal activity against Candida albicans. While Ismailia propolis had the highest antiviral activity against Reo-virus. Sharkia propolis showed the highest antibacterial activity against Staphylococcus aureus and moderate antiviral activity against infectious bursal disease virus and reovirus.     

Effect of Mexican propolis extracts from Apis mellifera on Candida albicans in vitro growth

Propolis is a resinous substance collected by bees (Apis mellifera) from different trees and bushes. Due to its antifungal, antibacterial, antiviral and antiparasitic properties, it has continued to be very popular throughout the time showing variable activity depending on its geographical origin. In Mexico, information about this product is very limited. The aim of this work was to evaluate the antifungal activity of four propolis ethanolic extracts from three different Mexican states, and four commercial extracts on Candida albicans growth. A reference strain (ATCC 10231) and 36 clinical isolates of C. albicans were used. The Minimal Inhibitory Concentration (MIC) was determined by the dilution on agar method. Growth curves on Sabouraud Dextrose broth with and without different propolis ethanolic extracts concentrations were performed. In addition, whether the effect was fungistatic or fungicide was determined. The propolis ethanolic extract obtained from Cuautitlán Izcalli, State of Mexico, showed the best biological activity, inhibiting 94.4% from the clinical isolates at 0.8 mg/ml; the reference strain was inhibited at 0.6 mg/ml. The propolis effect was fungistatic in low concentrations and fungicide in concentrations higher to MIC. The Mexican propolis ethanolic extract could be further investigated for its alternative use for the treatment of some C. albicans infections.     

In vitro activity of propolis against Streptococcus pyogenes

Propolis, a multifunctional substance used by bees to maintain the safety of their hives, is popular for its therapeutic potential against some micro-organisms. Ethanolic extracts of two propolis specimens, collected from different areas within a region in the north-west of Italy, were examined to evaluate their antimicrobial activity against 46 Streptococcus pyogenes strains. By both agar dilution and agar diffusion methods, the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were 相似文献   

Simple 1,4-benzoquinones with antibacterial activity from stems and leaves of Gunnera perpensa

From the dichloromethane extract of the leaves and stems of Gunnera perpensa two new, simple 1,4-benzoquinones and a known benzopyran-6-ol were isolated. From the methanol extract phytol was obtained. The two benzoquinones, 2-methyl-6-(-3-methyl-2-butenyl)benzo-1,4-quinone (1) and 3-hydroxy-2-methyl-5-(3-methyl-2-butenyl)benzo-1,4-quinone (2) and the benzopyran, 6-hydroxy-8-methyl-2,2-dimethyl-2H-benzopyran (3) were examined for antimicrobial properties together with the crude stem, leaf and root extracts. Minimum inhibitory concentration (MIC) assays were used to quantify antimicrobial activity and the MIC values for the crude extracts of stems, roots and leaves ranged between 100 microg and >16 mg/ml against the eight microorganisms investigated. Compound 1 showed significant antimicrobial activity with the most sensitive organism being Staphylococcus epidermidis with an MIC of 9.8 microg/ml. For compound 2, no activity was noted. Compound 3 exhibited good activity against the yeasts Cryptococcus neoformans (75 microg/ml) and Candida albicans (37.5 microg/ml).     

Antibacterial activity of honey and propolis from Apis mellifera and Tetragonisca angustula against Staphylococcus aureus

AIMS: The antibacterial activity against Staphylococcus aureus of honey and propolis produced by Apis mellifera and Tetragonisca angustula was evaluated. Secondary aims included the study of the chemical composition of propolis and honey samples and its relationship with antibacterial activity against S. aureus. METHODS AND RESULTS: The antibacterial activity of honey and propolis was determined by the method of macrodilution. The minimum inhibitory concentration (MICs) of A. mellifera honey ranged from 126.23 to 185.70 mg ml(-1) and of T. angustula from 142.87 to 214.33 mg ml(-1). For propolis, the MIC ranged from 0.36 to 3.65 mg ml(-1) (A. mellifera) and from 0.44 to 2.01 mg ml(-1) (T. angustula). Honey and propolis were evaluated by high-performance liquid chromatography. Some typical compounds of Brazilian propolis were also identified in honey samples. Principal component analysis revealed that the chemical composition of honey and propolis samples was distinct based on the geographical location of the samples. CONCLUSIONS: Propolis samples had higher antibacterial activity against S. aureus when compared with honey. However, both propolis and honey samples had antibacterial against S. aureus. SIGNIFICANCE AND IMPACT OF THE STUDY: These antimicrobial properties would warrant further studies on the clinical applications of propolis and honey against S. aureus.     

Antibacterial activity of the essential oil from Rosmarinus officinalis and its major components against oral pathogens

The essential oil of Rosmarinus officinalis L. (rosemary) was obtained by hydro-distillation and analysed by gas chromatography-mass spectrometry. Sixty-two constituents were identified, representing 98.06% of the total oil content. Oxygenated monoterpenes were the predominant components. The rosemary oil was characterized as having prominent (> 5%) contents of camphor (18.9%), verbenone (11.3%), a-pinene (9.6%), beta-myrcene (8.6%), 1,8-cineole (8.0%), and beta-caryophyllene (5.1%). The antimicrobial activity of the oil as well as of its major constituents was tested against the following microorganisms: Streptococcus mutans, Streptococcus mitis, Streptococcus sanguinis, Streptococcus salivarius, Streptococcus sobrinus, and Enterococcus faecalis, which are potentially responsible for the formation of dental caries in humans. The microdilution method was used for determination of the minimum inhibitory concentration (MIC) during evaluation of the antibacterial activity. The essential oil displayed low activity against the selected microorganisms. In the present study, the pure major compounds were more active than the essential oil. Among all the microorganisms tested, the pathogen S. mitis was the most susceptible and E. faecalis was the most resistant to the evaluated samples. This is the first report on antimicrobial activity of the major components of rosemary oil against oral pathogens.     

伊犁黑蜂蜂胶对不同状态下变形链球菌乳酸脱氢酶及其相关基因影响的实验研究

目的目的通过新疆伊犁黑蜂蜂胶乙醇提取物(Ethanol Extract of Propolis,EEP)对不同状态下变形链球菌乳酸脱氢酶活性及其相关基因表达影响的作用,研究伊犁黑蜂蜂胶抑制变形链球菌产酸的原因并探讨其可能的防龋机制。方法 (1)分别培养浮游状态与生物膜状态下生长的变形链球菌,根据实验分组用含梯度浓度EEP的BHI培养基、50 mg/L氟化钠的BHI培养基作用18 h,通过还原性辅酶I氧化法测定乳酸脱氢酶活性。(2)分别培养浮游状态与生物膜状态下生长变形链球菌,根据实验分组用含梯度浓度EEP的BHI培养基、含50 mg/L氟化钠的BHI培养基作用18 h,反转录-实时荧光定量PCR(RTq PCR)法测定各组乳酸脱氢酶编码基因ldh表达情况。结果 (1)在浮游状态与生物膜状态下,EEP组和Na F组乳酸脱氢酶活性均有降低,差异具有统计学意义(P0.05)。(2)浮游状态时,实验组组和阳性对照组ldh表达明显受到抑制(P0.05);生物膜状态下,实验组在1 MBEC、1/2 MBEC、1/4 MBEC浓度时ldh表达受到抑制(P0.05),Na F组ldh表达差异没有统计学意义(P0.05)。结论伊犁黑蜂蜂胶能够抑制浮游状态与生物膜状态下变形链球菌乳酸脱氢酶活性及其编码基因ldh表达,来抑制细菌产酸,伊犁黑蜂蜂胶可能是通过此途径抑制变形链球菌产酸,从而达到防龋的效果。     

Propolis: anti-Staphylococcus aureus activity and synergism with antimicrobial drugs

Propolis is a natural resinous substance collected by bees from tree exudates and secretions. Its antimicrobial activity has been investigated and inhibitory action on Staphylococcus aureus growth was evaluated. The in vitro synergism between ethanolic extract of propolis (EEP) and antimicrobial drugs by two susceptibility tests (Kirby and Bauer and E-Test) on 25 S. aureus strains was evaluated. Petri dishes with sub-inhibitory concentrations of EEP were incubated with 13 drugs using Kirby and Bauer method and synergism between EEP and five drugs [choramphenicol (CLO), gentamicin (GEN), netilmicin (NET), tetracycline (TET), and vancomycin (VAN)] was observed. Nine drugs were assayed by the E-test method and five of them exhibited a synergism [CLO, GEN, NET, TET, and clindamycin (CLI)]. The results demonstrated the synergism between EEP and antimicrobial drugs, especially those agents that interfere on bacterial protein synthesis.     

Antibacterial activity of triterpene acids and semi-synthetic derivatives against oral pathogens

Triterpene acids (ursolic, oleanoic, gypsogenic, and sumaresinolic acids) isolated from Miconia species, along with a mixture of ursolic and oleanolic acids and a mixture of maslinic and 2-a-hydroxyursolic acids, as well as ursolic acid derivatives were evaluated against the following microorganisms: Streptococcus mutans, Streptococcus mitis, Streptococcus sanguinis, Streptococcus salivarius, Streptococcus sobrinus, and Enterococcus faecalis, which are potentially responsible for the formation of dental caries in humans. The microdilution method was used for the determination of the minimum inhibitory concentration (MIC) during the evaluation of the antibacterial activity. All the isolated compounds, mixtures, and semi-synthetic derivatives displayed activity against all the tested bacteria, showing that they are promising antiplaque and anticaries agents. Ursolic and oleanolic acids displayed the most intense antibacterial effect, with MIC values ranging from 30 microg/mL to 80 microg/mL. The MIC values of ursolic acid derivatives, as well as those obtained for the mixture of ursolic and oleanolic acids showed that these compounds do not have higher antibacterial activity when compared with the activity observed with either ursolic acid or oleanolic acid alone. With regard to the structure-activity relationship of triterpene acids and derivatives, it is suggested that both hydroxy and carboxy groups present in the triterpenes are important for their antibacterial activity against oral pathogens.     

Potential application of Northern Argentine propolis to control some phytopathogenic bacteria

The antimicrobial activity of samples of Northern Argentine propolis (Tucumán, Santiago del Estero and Chaco) against phytopathogenic bacteria was assessed and the most active samples were identified. Minimal inhibitory concentration (MIC) values were determined by agar macrodilution and broth microdilution assays. Strong antibacterial activity was detected against Erwinia carotovora spp carotovora CECT 225, Pseudomonas syringae pvar tomato CECT 126, Pseudomonas corrugata CECT 124 and Xanthomonas campestris pvar vesicatoria CECT 792. The most active propolis extract (Tucumán, T1) was selected to bioguide isolation and identified for antimicrobial compound (2',4'-dihydroxychalcone). The antibacterial chalcone was more active than the propolis ethanolic extract (MIC values of 0.5-1 μg ml(-1) and 9.5-15 μg ml(-1), respectively). Phytotoxicity assays were realized and the propolis extracts did not retard germination of lettuce seeds or the growth of onion roots. Propolis solutions applied as sprays on tomato fruits infected with P. syringae reduced the severity of disease. Application of the Argentine propolis extracts diluted with water may be promising for the management of post harvest diseases of fruits.