The effect of Bulgarian propolis against Trypanosoma cruzi and during its interaction with host cells

Propolis has shown activity against pathogenic microorganisms that cause diseases in humans and animals. The ethanol (Et-Blg) and acetone (Ket-Blg) extracts from a Bulgarian propolis, with known chemical compositions, presented similar activity against tissue culture-derived amastigotes. The treatment of Trypanosoma cruzi-infected skeletal muscle cells with Et-Blg led to a decrease of infection and of the intracellular proliferation of amastigotes, while damage to the host cell was observed only at concentration 12.5 times higher than those affecting the parasite. Ultrastructural analysis of the effect of both extracts in epimastigotes revealed that the main targets were the mitochondrion and reservosomes. Et-Blg also affected the mitochondrion-kinetoplast complex in trypomastigotes, offering a potential target for chemotherapeutic agents.     

Activity of Brazilian and Bulgarian propolis against different species of Leishmania

Extracts of propolis samples collected in Brazil and Bulgaria were assayed against four Leishmania species--Leishmania amazonensis, L. braziliensis, L. chagasi from the New World, and L. major from the Old World--associated to different clinical forms of leishmaniasis. The composition of the extracts has been previously characterized by high temperature high resolution gas chromatography coupled to mass spectrometry. Considering the chemical differences among the extracts and the behavior of the parasites, it was observed significant differences in the leishmanicidal activities with IC50/1 day values in the range of 2.8 to 229.3 microg/ml . An overall analysis showed that for all the species evaluated, Bulgarian extracts were more active than the ethanol Brazilian extract. As the assayed propolis extracts have their chemical composition determined it merits further investigation the effect of individual components or their combinations on each Leishmania species.     

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.     

Biological activities from extracts of endophytic fungi isolated from Viguiera arenaria and Tithonia diversifolia

A total of 39 endophytic fungi have been isolated from Viguiera arenaria and Tithonia diversifolia, both collected in S?o Paulo State, Brazil. The isolates were identified based on their ribosomal DNA sequences. The ethyl acetate (EtOAc) extracts of all endophytic fungi were evaluated for their antimicrobial, antiparasitic and antitumoral activity. Antimicrobial screening was conducted using an agar diffusion assay against three pathogenic microorganisms: Staphylococcus aureus, Escherichia coli and Candida albicans. Antiparasitic activity was determined by enzymatic inhibition of gGAPDH of Trypanosoma cruzi and adenine phosphorybosiltransferase (APRT) of Leishmania tarentolae. Antitumoral activity was tested against human T leukemia cells by the Mosmann colorimetric method. All extracts showed activity in at least one assay: 79.5% of the extracts were cytotoxic against leukemia cells, 5.1% of the extracts were active against S. aureus, 25.6% against E. coli and 64.1% against Candida albicans. Only one extract showed promising results in the inhibition of parasitic enzymes gGAPDH (95.0%) and three were found to inhibit APRT activity. The cytotoxic extract produced by the strain VA1 (Glomerella cingulata) was fractionated and yielded nectriapyrone and tyrosol. Nectriapyrone showed relevant cytotoxic activity against both human T leukemia and melanoma tumor cell lines.     

Antibacterial and free-radical scavenging activities of Sonoran propolis

AIMS: To evaluate the antibacterial and free-radical scavenging (FRS) activities of propolis collected from three different areas of Sonoran Desert in northwestern Mexico [Pueblo de Alamos (PAP), Ures (UP) and Caborca (CP)]. METHODS AND RESULTS: The antibacterial and FRS activities of Sonoran propolis were determined by the broth microdilution method and the DPPH (1,1-diphenyl-2-picrylhydracyl) assay, respectively. Propolis samples had antibacterial activity against only Gram-positive bacteria. The UP sample showed the highest antibacterial activity against Staphylococcus aureus [minimal inhibitory concentration (MIC) 100 microg ml(-1)] in a concentration-dependent manner (UP > CP > PAP). Caffeic acid phenethyl ester (CAPE), a UP propolis constituent, had very high growth-inhibitory activity towards Gram-positive bacteria, particularly against S. aureus (MIC 0.1 mmol l(-1)). To our knowledge, this is the first study showing a strong antibacterial activity of CAPE against S. aureus. Additionally, propolis CP exhibited high FRS activity (86% +/- 0.3 at 100 microg ml(-1)) comparable with those of the reference antioxidants vitamin C (87.4% +/- 1.7 at 70 micromol l(-1)) and BHT (66.07% +/- 0.76 at 140 micromol l(-1)). The propolis compounds CAPE and rutin showed high FRS activity (90.4% +/- 0.2 and 88.5% +/- 0.8 at 70 micromol l(-1), respectively). CONCLUSIONS: Sonoran propolis UP and CAPE had strong antibacterial activity against S. aureus. In addition, propolis CP showed potent FRS activity comparable with those of vitamin C and BHT. SIGNIFICANCE AND IMPACT OF THE STUDY: The strong antibacterial and antioxidant properties of Sonoran propolis and some of its constituents support further studies on the clinical applications of this natural bee product against S. aureus and several oxidative damage-related diseases.     

Trichophyton species susceptibility to green and red propolis from Brazil

Aims:  The in vitro antifungal activity of Brazilian green and red propolis was tested against different species of Trichophyton .
Methods and Results:  The antifungal activity of the Brazilian aqueous and alcoholic extracts of the green propolis and the alcoholic extract of red propolis was observed against Trichophyton rubrum , Trichophyton tonsurans and Trichophyton mentagrohytes samples, using as controls itraconazole and terbinafine. The minimal inhibitory concentration was determined following the microdilution method indicated by the 'Clinical and Laboratory Standards Institute'. The minimal fungicide concentration was determined by the absence of growth in liquid sabouraud culture medium. The data obtained showed that the green propolis alcoholic extract's antifungal activity was from 64 to 1024 μg ml⁻¹, whereas the red propolis alcoholic extract was from 8 to 1024 μg ml⁻¹.
Conclusions:  The antifungal activity of the red propolis alcoholic extract was more efficient than the green propolis alcoholic extract for all three species studied. The T. rubrum samples were shown to be more sensitive to the antifungal activity of the alcoholic extracts of the propolis.
Significance and Impact of the Study:  The antifungal potential of the alcoholic extracts of green and red propolis demonstrated suggest an applicable potential as an alternative treatment for dermatophytosis caused by these species.     

HPTLC-MS as an efficient hyphenated technique for the rapid identification of antimicrobial compounds from propolis

The well-known anti-infective properties of propolis are determined by its chemical composition, which in turn is influenced by geographical factors and reflects the botanical diversity in the vicinity of the beehive. Although there are several reports on the anti-infective properties of crude propolis, few are aimed at identifying specific compound(s) responsible for the observed activities. Using South African propolis as an example, the application of high performance thin layer chromatography-bioautography in tandem with mass spectrometry was investigated for the rapid identification of antimicrobial and anti-quorum sensing (anti-QS) compounds. Pinocembrin was found to be responsible for the observed antifungal activity of the propolis against Candida albicans. Three compounds were found to be active against all of the evaluated Gram-positive and Gram-negative bacteria. The identity of the first was confirmed as pinobanksin, one remains unidentified, while the third corresponds to either pinobanksin 3-O-pentanoate or 2-methylbutyrate. The identification of caffeic acid as the anti-QS component was confirmed quantitatively using the violacein inhibitory assay.     

Antimicrobial activity of isocytisoside and extracts of Aquilegia vulgaris L

AIMS: The aim of this study was to analyse the antimicrobial properties extracts of Aquilegia vulgaris, and their principial flavonoid component and to compare the obtained results with the activity of gentamicin and nystatin. METHODS AND RESULTS: The ethanol, acetone and isopropanol extracts as well as the subextracts isolated from the methanol extract together with the main flavonoid: 4'-methoxy-5,7-dihydroxyflavone 6-C-glucoside (isocytisoside) were obtained from the leaves with stems of Aquilegia vulgaris L. All the extracts were analysed by TLC to confirm flavonoids and phenolic acids occurrence. The antimicrobial activity was tested by the method of series dilutions against different Gram-positive, Gram-negative bacteria and also fungi. The results have shown that the extracts, subextracts and isocytisoside inhibit growth of all studied micro-organisms, revealing the greatest activity against Gram-positive Staphylococcus aureus, Staph. epidermidis and the mould Aspergillus niger. CONCLUSIONS: The antimicrobial activity of the tested materials it is possibly related to the content of isocytisoside. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has determined new activity of A. vulgaris and suggested the necessity of further studies.     

In vitro antimicrobial activity of a novel propolis formulation (Actichelated propolis)

AIMS: This study compared in vitro activities of Actichelated propolis (a multicomposite material obtained with mechano-chemichal activation) and of a hydroalcoholic extract of propolis. METHODS AND RESULTS: Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC), determined by means of microdilution broth method, against five strains of Staphylococcus aureus, Streptococcus pyogenes, Haemophilus influenzae, Enterococcus spp., Escherichia coli, Proteus mirabilis and Pseudomonas aeruginosa, showed a greater potency of Actichelated propolis (MIC range: 0.016-4 mg flavonoids ml(-1)) in respect to the hydroalcoholic extract (MIC range: 0.08-21.4 mg flavonoids ml(-1)). Concentrations of Actichelated propolis active against adenovirus, influenza virus, parainfluenza virus and herpes virus type 1 were at least 10 times lower than those of the hydroalcoholic extract. Preincubation of Strep. pyogenes and H. influenzae with subinhibitory concentrations of Actichelated propolis (1/4 and 1/8 x MIC) significantly reduced the number of bacteria that adhered to human buccal cells. CONCLUSIONS: Actichelated propolis has proven to possess antibacterial and antiviral activity higher than a hydroalcoholic extract, being also able to interfere on bacterial adhesion to human oral cells. SIGNIFICANCE AND IMPACT OF THE STUDY: This new formulation of propolis showing better antimicrobial and physical characteristics could improve the application of propolis in respiratory tract infections.     

Comparative study of in vitro methods used to analyse the activity of propolis extracts with different compositions against species of Candida

AIMS: Propolis is known for its activity against micro-organisms and different in vitro assays have been used to evaluate this activity, frequently with contradictory results. METHODS AND RESULTS: Brazilian propolis from the state of S?o Paulo was extracted by maceration using different concentrations of ethanol and water. The resultant extracts were analysed by chromatographic methods. Several microbiological methods were compared to determine which one best evaluated the activity of the propolis extracts against species of Candida, with average minimal inhibitory concentration values between 6 and 12 mg ml(-1). CONCLUSIONS: Agar dilution in plates showed the clearest results. These were in agreement with the chromatographic analyses, which also identified the active substances. SIGNIFICANCE AND IMPACT OF THE STUDY: Although the active substances identified in this sample are typical of Brazilian propolis, their activity against Candida had not been recognized previously, demonstrating the importance of standardizing the correct combination of microbiological and chromatographic analyses.     

Chemical compositions and antimicrobial activities of four different Anatolian propolis samples

Propolis means a gum that is gathered by bees from various plants. It is known for its biological properties, having antibacterial, antifungal and healing properties. The aims of this study were to evaluate the antimicrobial activity of four different Anatolian propolis samples on different groups of microorganisms including some oral pathogens and comparison between their chemical compositions. Ethanol extracts of propolis (EEP) were prepared from four different Anatolian propolis samples and examined whether EEP inhibit the growth of the test microorganisms or not. For the antimicrobial activity assays, minimum inhibitory concentrations (MIC) were determined by using macrodilution method. The MIC values of the most effective propolis (TB) were 2 microg/ml for Streptococcus sobrinus and Enterococcus faecalis, 4 microg/ml for Micrococcus luteus, Candida albicans and C. krusei, 8 microg/ml for Streptococcus mutans, Staphylococcus aureus, Staphylococcus epidermidis and Enterobacter aerogenes, 16 microg/ml for Escherichia coli and C. tropicalis and 32 microg/ml for Salmonella typhimurium and Pseudomonas aeruginosa. The chemical compositions of EEP's were determined by high-temperature high-resolution gas chromatography coupled to mass spectrometry. The main compounds of four Anatolian propolis samples were flavonoids such as pinocembrin, pinostropin, isalpinin, pinobanksin, quercetin, naringenin, galangine and chrysin. Although propolis samples were collected from different regions of Anatolia all showed significant antimicrobial activity against the Gram positive bacteria and yeasts. Propolis can prevent dental caries since it demonstrated significant antimicrobial activity against the microorganisms such as Streptococcus mutans, Streptococcus sobrinus and C. albicans, which involves in oral diseases.     

Propolis standardized extract (EPP-AF®), an innovative chemically and biologically reproducible pharmaceutical compound for treating wounds

The aim of this study was to develop a formulation, containing the propolis standardized extract (EPP-AF^®), which can assist in the healing of skin lesions. To achieve this objective the antimicrobial activity and chemical composition of the propolis extract was determined. The final product was subjected to in vitro and in vivo pre-clinical evaluation. The broth macrodilution method was used to determine the antimicrobial activity of the extracts and formulations against the microorganisms most commonly found in burns, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus and Staphylococcus epidermidis. Wistar rats with puncture wounded skin were used to evaluate the wound healing properties of propolis. The results of chemical and biological characterization demonstrated the batch-to-batch reproducibility of the standardized extract which is an unprecedented result. The antimicrobial and wound healing activity of the pharmaceutical studied showed the best results when samples contain 3.6% propolis, suggesting that this is the most promising composition.     

Chemical composition and biological activity of propolis from Brazilian meliponinae

Twenty-one propolis samples produced by 12 different Meliponinae species were analyzed by GC-MS. Several chemical types of stingless bees' propolis could be grouped, according to the prevailing type of compounds like: 'gallic acid", "diterpenic" and "triterpenic" types. The results confirm that neither the bee species nor the geographical location determine the chemical composition of Meliponinae propolis and the choice of its plant source, respectively. This could be explained by the fact that Meliponinae forage over short distances (maximum 500 m) and thus use as propolis source the first plant exudate they encounter during their flights. The antibacterial, antifungal and cytotoxic activities of the samples were also investigated. Most samples had weak or no activity against E. coli, weak action against Candida albicans. Some of them showed significant activity against St. aureus., presumably connected to the high concentration of diterpenic acids. Samples rich in diterpenic acids possessed also high cytotoxic activity (Artemia salina test).     

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.     

Chemical properties and antioxidant and antimicrobial activities of slovenian propolis

The chemical composition as well as the antioxidant and antimicrobial activities of two EtOH extracts of propolis (PEEs) from Slovenia were determined. EtOH was used as extracting solvent at 70 and 96%, providing the extracts PEE70 and PEE96, respectively. The extraction with 70% EtOH was more efficient than that with 96% EtOH, as the PEE70 was richer in total phenolic compounds than the PEE96. The Slovenian propolis was characterized by different phenolic acids and flavonoids. The PEE96 was slightly richer in three specific compounds, i.e., caffeic acid, ferulic acid, and luteolin, while all other substances detected showed higher contents in the PEE70. The PEE70 showed a stronger reducing power and ability to scavenge free radicals and metal ions than the PEE96. Both PEEs were in the main more effective against Gram-positive bacteria than against fungi and Gram-negative bacteria like Salmonella and Escherichia coli, with the exception of Campylobacter. The PEE96 decreased the intracellular oxidation in Saccharomyces cerevisiae in a dose-dependent manner. The antimicrobial activities and antioxidant properties were related to the total phenolic contents. The two PEEs have the potential for use as natural antimicrobial and antioxidant additives in foods.     

Antimicrobial activity of extracts of chemical races of the lichen Pseudevernia furfuracea and their physodic acid, chloroatranorin, atranorin, and olivetoric acid constituents

The antimicrobial activity and the MIC values of the ethanol, chloroform, diethyl ether, and acetone extracts of the chemical races of Pseudevernia furfuracea (var. furfuracea and var. ceratea) and their physodic acid, chloroatranorin, atranorin, and olivetoric acid constituents have been investigated against some microorganisms. Nearly all extracts of both chemical races showed antimicrobial activity against Aeromonas hydrophila, Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Proteus vulgaris, Staphylococcus aureus, Streptococcus faecalis, Yersinia enterocolitica, Candida albicans, Candida glabrata, Alternaria alternata, Ascochyta rabiei, Aspergillus niger, Fusarium culmorum, Fusarium moniliforme, Fusarium oxysporum, Fusarium solani, and Penicillium notatum. There was no antimicrobial activity of the extracts against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Pseudomonas syringae, Salmonella typhimurium, Alternaria citri, Alternaria tenuissima, and Gaeumannomyces graminis. Chloroatranorin and olivetoric acid were active against the same microorganisms with few exceptions. Physodic acid was active against about the same bacteria and yeasts and inactive against all of the filamentous fungi tested. Also no activity of atranorin against the filamentous fungi was observed.     

Terpenes with antimicrobial activity from Cretan propolis

Five terpenes, the diterpenes: 14,15-dinor-13-oxo-8(17)-labden-19-oic acid and a mixture of labda-8(17),13E-dien-19-carboxy-15-yl oleate and palmitate as well as the triterpenes, 3,4-seco-cycloart-12-hydroxy-4(28),24-dien-3-oic acid and cycloart-3,7-dihydroxy-24-en-28-oic acid were isolated from Cretan propolis. Moreover, 18 known compounds were also isolated, seven of them for the first time as propolis components. All structures were established on the basis of spectroscopic analysis and chemical evidence. All isolated compounds were tested for antimicrobial activity against some Gram-positive and Gram-negative bacteria as well as against some human pathogenic fungi showing a broad spectrum of antimicrobial activity.     

伊犁黑蜂蜂胶对不同生长状态白假丝酵母菌抑菌作用的体外研究

目的探究伊犁黑蜂蜂胶对白假丝酵母菌游离状态及其生物膜状态的影响。方法采用NCCLS M27-A2酵母微量稀释法结合平板法测定蜂胶对游离状态下白假丝酵母菌的最小抑菌浓度(MIC)和最低杀菌浓度(MBC);玻片法在体外构建白假丝酵母菌生物膜模型,采用MTT法检测蜂胶对白假丝酵母菌生物膜状态下的最低生物膜清除浓度(MBEC)及抑制作用;激光共聚焦显微镜(CLSM)观察不同浓度蜂胶醇提取物对同一时间白假丝酵母菌生物膜的抑菌效果,并进行红蓝荧光染色定量分析。结果伊犁黑蜂蜂胶对浮游白假丝酵母菌的MIC为4mg/mL,MBC为8mg/mL;伊犁黑蜂蜂胶对生物膜状态下白假丝酵母菌的MBEC为16mg/mL,随着药物浓度的增加抑菌性逐渐增大,差异具有统计学意义(P0.05);CLSM观察,伊犁黑蜂蜂胶作用于白假丝酵母菌生物膜后可使生物膜内活菌比例降低,生物膜活性减弱,生物膜内实验菌死亡率随蜂胶浓度增加呈上升趋势,与阴性对照组比较差异有统计学意义(P0.05)。结论伊犁黑蜂蜂胶在体外对不同状态下的白假丝酵母菌均起到了明显的抑制作用。     

Antifungal activity of Apulia region propolis

A study was carried out to assess the in vitro antifungal activity of some natural Apulian propolis extracts of different origin. Their antifungal activity was compared to the antifungal activity of conifers and commercial propolis extracts. All extracts revealed antifungal activity against dermatophytes and Candida species. The antifungal activity differences found depended on the origin of the propolis and the solvent used for extraction. The best antifungal activity was given by the 'Orimini' propolis. The antifungal activity may have been influenced by the presence of different cinnamic and flavonoid components and their different concentration in the extracts. Further investigations are needed to validate this hypothesis.     

In vitro activity of Bulgarian propolis against 94 clinical isolates of anaerobic bacteria

The aim was to evaluate the effect of 30% ethanolic extract of Bulgarian propolis on 94 clinical anaerobic strains. The strains were tested by both agar-well diffusion (wells, 7 mm diameter) and disk-diffusion methods. Only 15% of Clostridium-, 3.3% of other Gram-positive- and 9.1% of Gram-negative anaerobic strains were not inhibited by 30 microL propolis extract per well. Propolis extract was more active than the ethanol (P < 0.001). By 30 microL extract per well, mean inhibitory diameters of the clostridia, other Gram-positive- and Gram-negative anaerobes were 11.5, 13.1, and 11.3 mm, and those by 90 microL were 16, 18.1 and 15.4 mm, respectively. Mean inhibitory diameters of all strains by 30 and 90 microL ethanol were only 8.4 and 9.5 mm. By 30 microL propolis extract per well, inhibitory diameters of 15 mm or more were more common in Gram-positive (32%) than in Gram-negative bacteria (13.6%, P < 0.05). Moist propolis disks inhibited more strains (89.4%) than dried disks (68.1%, P < 0.001). Most (81.8%) Bacteroides fragilis group strains and 75% of clostridial strains were inhibited by moist EEP disks. CONCLUSION: Bulgarian propolis was active against most anaerobic strains of different genera. In addition to oral pathogens, an activity of propolis against Clostridium, Bacteroides and Propionibacterium species was observed. The results could motivate a higher medical interest and further trials for evaluating the use of bee glue for prophylaxis or treatment of some anaerobic infections such as oral, skin and wound diseases.