Propolis from native Stingless Bees: ultrasound-assisted extraction / Propóleos de abejas nativas sin aguijón: extracción asistida por ultrasonido

BACKGROUND: Propolis has been considered a highly valuable material due to its therapeutic properties. However, in Colombia, the commercialization of propolis is limited not only by low production but also by the little knowledge about its efficient extraction. Therefore, finding an optimal and economical extraction method to obtain propolis is a necessity for beekeepers that would open new possibilities for industrial use and, therefore, for the market. OBJECTIVES:The objective of this study was to evaluate a conventional and ultrasound-assisted extraction method, seeking to obtain the highest yield and a high amount of content of bioactive compounds in propolis extracts. METHODS: The extraction was carried out for three crude propolis from different types of bees: Tetragoniscaangustula or Angelita (ANG), Meliponaeburnea or Melipona(MEL), and Scaptotrigonaspp (SCT). The extracts were characterized by color, pH, visual appearance, solid content, antioxidant capacity, total polyphenol content, and bacterial inhibition capacity. RESULTS: The highest extraction performance was obtained when the ultrasound-assisted method was used, especially for the ANG extract, which in addition to presenting inhibition for gram-negative (E. coli) and gram-positive (S. Aureus) bacteria, had the best antioxidant activity with a value of 545 mg GAE / 100 g of sample and total polyphenol content of 1,884 mg GAE / 100 g of sample. CONCLUSIONS: Ultrasound-assisted extraction can be considered a low-cost alternative to increase the extraction performance of crude propolis, together with its total polyphenol content and antioxidant capacity, without altering its physical properties

ANTECEDENTES: El propóleos ha sido considerado un material de alto valor por sus propiedades terapéuticas. Sin embargo, en Colombia la comercialización de propóleos está limitada no solo por la baja producción sino también por el incipiente conocimiento sobre la extracción eficiente de este. Por ello, encontrar un método de extracción óptimo y económico para la obtención de propóleos es una necesidad para los apicultores que abriría nuevas posibilidades para el uso industrial y por tanto para el mercado. OBJETIVOS: El objetivo de este estudio fue evaluar un método de extracción convencional y asistido por ultrasonido (US) buscando el mayor rendimiento y alto contenido de compuestos bioactivos en extractos de propóleos. MÉTODOS: La extracción se realizó para tres propóleos crudos de diferentes tipos de abejas Tetragonisca angustula o Angelita(ANG), Melipona eburnea o Melipona (MEL) y Scaptotrigona spp (SCT). Todos los extractos se caracterizaron por su color, pH, apariencia visual, contenido de sólidos, capacidad antioxidante, contenido total de polifenoles y capacidad de inhibición bacteriana. RESULTADOS: El mayor rendimiento de extracción se obtuvo cuando se usó el método asistido por ultrasonido y específicamente para el extracto ANG, que además de presentar inhibición para bacterias gram negativas (E. coli) y gram positivas (S. Aureus), tuvo la mejor actividad antioxidante con un valor de 545 mg GAE / 100 g de muestra y contenido total de polifenoles de 1884 mg GAE / 100 g de muestra. CONCLUSIONES: La extracción asistida por ultrasonido puede considerarse una alternativa de bajo costo para aumentar el rendimiento de extracción del propóleos crudo, así como su contenido total de polifenoles y capacidad antioxidante sin alterar sus propiedades físicas

Brazilian Green Propolis: Chemical Composition of Essential Oil and Their In Vitro Antioxidant, Antibacterial and Antiproliferative Activities

Abstract Propolis is a resinous substance collected and processed by Apis mellifera from parts of plants, buds and exudates. In Minas Gerais (MG) state, Brazil, green propolis is produced from the collection of resinous substance found in shoot apices of Baccharis dracunculifolia. This paper aims to investigate the chemical composition and in vitro antioxidant, anti-Helicobacter pylori, antimycobacterial and antiproliferative activities of essential oil (EO) from Brazilian green propolis (BGP-EO). The oil showed high antibacterial activity against H. pylori (MIC = 6.25 µg/mL), Mycobacterium avium (MIC = 62.5 µg/mL) and M. tuberculosis (MIC = 64 µg/mL). Its antioxidant activity was evaluated in vitro by both DPPH (IC50 = 23.48 µg/mL) and ABTS (IC50 = 32.18 µg/mL) methods. The antiproliferative activity in normal (GM07492A, lung fibroblasts) and tumor cell lines (MCF-7, HeLa and M059J) was analyzed by the XTT assay. BGP-EO showed inhibition of normal cell growth at 68.93 ± 2.56 µg/mL. Antiproliferative activity was observed against human tumor cell lines, whose IC50 values were 56.17, 66.43 and -65.83 µg/mL for MCF-7, HeLa and M059J cells, respectively. Its major constituents, which were determined by GC-FID and GC-MS, were carvacrol (20.7 %), acetophenone (13.5 %), spathulenol (11.0 %), (E)-nerolidol (9.7 %) and β-caryophyllene (6.2 %). These results showed the effectiveness of BGP-EO as a natural product which has promising biological activities.

Phytochemical screening and in vitro antibacterial, antifungal, antioxidant and antitumor activities of the red propolis Alagoas / Triagem fitoquimico e atividade antibacteriana, antifúngica, antioxidante e antitumoral in vitro da propólis vermelha de Alagoas

Abstract The study aimed to evaluate the antimicrobial activity, antioxidant, toxicity and phytochemical screening of the Red Propolis Alagoas. Antimicrobial activity was evaluated by disk diffusion method. Determination of antioxidant activity was performed using the DPPH assay (1.1-diphenyl-2-picrylhydrazyl), FTC (ferric thiocyanate) and determination of phenolic compounds by Follin method. Toxicity was performed by the method of Artemia salina and cytotoxicity by MTT method. The phytochemical screening for the detection of allelochemicals was performed. The ethanol extract of propolis of Alagoas showed significant results for antimicrobial activity, and inhibitory activity for Staphylococcus aureus and Candida krusei. The antioxidant activity of the FTC method was 80% to 108.3% hydrogen peroxide kidnapping, the DPPH method showed an EC50 3.97 mg/mL, the content of total phenolic compounds was determined by calibration curve gallic acid, resulting from 0.0005 mg/100 g of gallic acid equivalent. The extract was non-toxic by A. salina method. The propolis extract showed high activity with a higher percentage than 75% inhibition of tumor cells OVCAR-8, SF-295 and HCT116. Chemical constituents were observed as flavonones, xanthones, flavonols, and Chalcones Auronas, Catechins and leucoanthocyanidins. It is concluded that the extract can be tested is considered a potential source of bioactive metabolites.

Resumo O trabalho teve como objetivo avaliar a atividade antimicrobiana, antioxidante, a toxicidade e a prospecção fitoquímica da Própolis Vermelha de Alagoas. A atividade antimicrobiana foi avaliada pelo método de difusão em disco. A determinação do potencial antioxidante foi realizada utilizando o método de DPPH (1,1-difenil-2-picrilhidrazil), FTC (Tiocianato Férrico) e determinação de compostos fenólicos pelo método de Follin. A toxicidade foi realizada pelo método de Artemia salina e a citotoxicidade pelo método do MTT. Foi realizada a prospecção fitoquímica para a pesquisa de aleloquímicos. O extrato etanólico da própolis vermelha de Alagoas apresentou resultados significantes para atividade antimicrobiana, tendo a atividade inibitória para Staphylococcus aureus e Candida krusei. Quanto a atividade antioxidante o método de FTC teve 80% a 108,3% de sequestro de peróxido de hidrogênio, o método de DPPH apresentou um CE50 de 3,97 μg/mL, o teor de compostos fenólicos totais foi determinado mediante curva de calibração do ácido gálico, tendo resultado de 0,0005 mg/100 g equivalente de ácido gálico. O extrato foi atóxico pelo método de A. salina. O extrato da própolis mostrou elevada atividade com percentual de inibição maior que 75% sobre células tumorais OVCAR-8, SF-295 e HCT116. Foram observados constituintes químicos como flavononas, xantonas, flavonóis, Chalconas e Auronas, Catequinas e Leucoantocianidinas. Conclui-se que o extrato testado pode ser considerado é uma fonte potencial de metabólitos bioativos.

Evaluación in vitro de la actividad antimicrobiana de un propóleo chileno sobre muestras clínicas de exudados bucofaríngeos y cepas ATCC / In vitro evaluation of the antimicrobial activity of a Chilean propolis on clinical samples of oropharyngeal exudates

Propolis is a substance manufactured by Apis mellifera and has been widely used in folk medicine due to its high concentration of bioactive compounds. The purpose of the following study was to characterize and evaluate in vitro the antimicrobial properties of propolis on clinical samples and ATCC strains. The chemical characterization of propolis presents a concentration of total polyphenols of 247 ± 9 mg EAG g-1 MS, flavones and flavonols 75± 4 mg EQ g-1 MS, flavanonones and flavanonols 118 ± 11 EP g-1 MS. HPLC-DAD identified apigenin, galangin, phenethyl ester of caffeic acid and pinocembrin, in addition to 16 compounds by HPLC MS/MS. Chilean propolis is a natural antimicrobial, showing effectiveness in strains ATCC Staphylococcus aureus, Candida albicans, Trichophyton rubrum and clinical samples of Staphylococcus aureus unlike Escherichia coli. These results demonstrate the antimicrobial effectiveness of the synergy of compounds present in propolis against different human pathogens.

El propóleos es una substancia fabricada por Apis mellifera y ha sido utilizado ampliamente en la medicina popular debido a su alta concentración de compuestos bioactivos. El propósito del siguiente estudio fue caracterizar y evaluar in vitro las propiedades antimicrobianas del propóleos sobre muestras clínicas y cepas ATCC. La caracterización química de propóleos presenta una concentración de polifenoles totales de 247 ± 9 mg EAG g-1 de MS, flavonas y flavonoles 75 ± 4 mg EQ g-1 de MS, flavanononas y flavanonoles 118 ± 11 EP g-1 de MS. Mediante HPLC-DAD se identificó apigenina, galangina, fenetil éster del ácido cafeico y pinocembrina, además de 16 compuestos mediante HPLC MS/MS. El propóleos chileno es un antimicrobiano natural, observándose efectividad en cepas ATCC Staphylococcus aureus, Candida albicans, Trichophyton rubrum y muestras clínicas de Staphylococcus aureus a diferencia de Escherichia coli. Estos resultados demuestran la efectividad antimicrobiana de la sinergia de compuestos presentes en el propóleos ante diferentes patógenos humanos.

Evaluación de la actividad anti-inflamatoria de propóleos chileno sobre cortes histológicos de orejas de ratón / Evaluation of the anti-inflammatory activity of Chilean propolis on histological sections of mouse ears

RESUMEN: El propóleos es un producto resinoso complejo producido por las abejas Apis mellifera, el cual posee diversas actividades biológicas como inmunomodulador, antiinflamatorio, anticancerígeno, antiviral, antibacteriano, antioxidante, entre otros. El propósito del siguiente estudio fue realizar una evaluación in vivo de las propiedades antiinflamatorias de un extracto de propóleos chileno, sobre el modelo de edema auricular inducido por 13-acetato-12-O-tetradecanoilforbol (TPA) en pabellón auricular de ratón, para posterior evaluación y análisis histológico. El extracto de propóleos chileno (EEP) utilizado se obtuvo a partir de un macerado etanólico, rotaevaporado y liofilizado. Se observó que el EEP disminuyó el edema y el infiltrado inflamatorio de forma significativa. Estos resultados sugieren que el extracto etanólico de propóleos chileno posee potenciales efectos antiinflamatorios o moduladores del sistema inmunológico en edema auricular.

SUMMARY: Propolis is a complex resinous product produced by bees Apis mellifera, which has a number of biological activities such as an immunomodulator, anti-inflammatory, anticarcinogenic, antiviral, antibacterial, antioxidant, among others. The purpose of the following study was to perform an in vivo evaluation of the anti-inflammatory properties of a Chilean propolis extract, on the model of atrial edema induced 12-O-tetradecanoyl phorbol-13- acetate (TPA) in the mouse auricular pavilion, for later evaluation and histological analysis. The Chilean propolis extract (EPP) used was obtained from an ethanolic, rotaevaporated and lyophilized macerate. It was observed that the EPP significantly decreased edema and inflammatory infiltrate. These results suggest that the ethanolic extract of Chilean propolis possesses potential anti-inflammatory or modulatory effects of the immune system in atrial edema.

Chemical characterization, antioxidant and antimicrobial activity of propolis obtained from Melipona quadrifasciata quadrifasciata and Tetragonisca angustula stingless bees

In this study, we investigated the chemical composition, and antioxidant and antibacterial properties of ethanolic extracts of propolis (EEP) from Melipona quadrifasciata quadrifasciata and Tetragonisca angustula. Chemical composition of EEP was determined by colorimetry and chromatographic (HPLC-DAD and UPLC-Q/TOF-MS/MS) analysis. Antimicrobial activity of EEP was evaluated against gram-positive (S. aureus, methicillin-resistant S. aureus, E. faecalis) and gram-negative (E. coli and K. pneumoniae) bacteria by the minimal inhibitory concentration (MIC) test using the microdilution method. Furthermore, the growth curve and integrity of cell membrane of S. aureus and E. coli were investigated using standard microbiological methods. HPLC-DAD analysis showed that the EEP of M. quadrifasciata quadrifasciata has a more complex chemical composition than the EEP of T. angustula. Moreover, UPLC-MS analyses of M. quadrifasciata quadrifascita indicated flavonoids and terpenes as major constituents. The bactericidal activity of both EEPs was higher against gram-positive bacteria than for gram-negative bacteria. The EEP from M. quadrifasciata quadrifasciata presented MIC values lower than the EEP from T. angustula for all tested bacteria. The EEP from M. quadrifasciata quadrifasciata caused lysis of the bacterial wall and release of intracellular components from both E. coli and S. aureus. Our findings indicate that the chemical composition of propolis from stingless bees is complex and depends on the species. The extract from M. quadrifasciata quadrifascita was more effective against gram-positive than gram-negative strains, especially against S. aureus and methicillin-resistant S. aureus compared to T. angustula extract, by a mechanism that involves disturbance of the bacterial cell membrane integrity.

Susceptibilidad de Cepas de Candida Oral a Extracto Etanólico del Propóleo Chileno de Olmué / Susceptibility of Oral Candida Strains to Ethanol Extract of Chilean Propolis of Olmué

RESUMEN: La infección por Candida albicans en la mucosa oral es conocida como Candidiasis oral (CO) y se diagnostica por el reconocimiento de cambios clínicos y la presencia de pseudohifas, hifas o levaduras en muestras obtenidas por citología exfoliativa o biopsia. Los agentes farmacológicos tópicos clásicos para el tratamiento de CO son Nistatina y Miconazol. Sin embargo, a pesar de las distintas terapias contra CO, existen formas de Candida resistentes al tratamiento convencional. El objetivo de este estudio fue determinar la susceptibilidad in vitro de Candida spp. a un extracto etanólico de propóleo de Olmué. Se realizó un estudio experimental descriptivo in vitro en donde se evaluó el efecto que presenta el uso de extracto etanólico de propóleo como antifúngico sobre cepas de Candida spp. obtenidas de la cavidad oral (mucosa palatina) de 31 individuos, con candidiasis oral diagnosticados con estomatitis subprotésica. El propóleo chileno utilizado fue obtenido de la zona geográfica de Olmué, quinta región. Se encontró que el 100 % de las muestras en rangos de concentración de propóleo de 0,1 µg/mL y 1,6 µg/mL presentaron un grado de inhibición en el crecimiento de Candida Oral y por otra parte el extracto etanólico de propóleo que generó inhibición en la mayor cantidad de muestras fue al 0,4 µg/mL (41,94 % de las muestras) y en segundo lugar la concentración al 0,2 µg/mL (35 % de las muestras). Se concluyó que el extracto etanólico de propóleo chileno obtenido de la zona de Olmué presenta la capacidad de inhibir el crecimiento de Candida spp. en agar Sabouraud in vitro de forma dosis dependiente.

ABSTRACT: Fungal (or yeast) infections; mycoses, occurring in the oral mucous membranes, of Candida species (mostly C. albicans, a normal component of the oral microbiota), also known as oral thrush or oral candidiasis (OC), can be diagnosed via the recognition of clinical changes and the presence of pseudohyphae, hyphae or yeasts in samples obtained by exfoliative cytology and/or biopsy. Topical pharmacological preparations and drugs such as Nystatin and Miconazole are used in the treatment of CO. However, there are forms of Candida with resistance to such conventional treatment approaches. Therefore, the aim of this study is to determine the in vitro susceptibility of Candida spp.; an ethanolic extract of propolis from Olmué. Hence, an experimental in vitro descriptive study was carried out in which the effect of an ethanolic extract of propolis used as antifungal on strains of Candida spp. obtained from the oral cavity (palatine mucosa) of 31 individuals, diagnosed with OC (subdenture stomatitis) is determined. Natural propolis was obtained from the Olmué area, in the 5th region of Chile. It was found that 100 % of the samples with propolis concentration ranging from 0.1 mg / mL to 1.6 mg / mL presented a degree of inhibition in the growth of OC. On the other hand, the ethanolic extract of propolis that generated inhibition in the largestnumber of samples was 0.4 mg / mL (41.94 % of the samples) followed by the concentration of 0.2 mg / mL (35 % of the samples). Therefore, it can be concluded that the ethanolic extract of Chilean propolis obtained from the Olmué area has the ability to inhibit the growth of Candida spp. in vitro in a dosage-dependent manner.

Propolis from Southwest of Parana produced by selected bees: Influence of seasonality and food supplementation on antioxidant activity and phenolic profile

ABSTRACT Propolis produced by selected bees Apis mellifera were collected from March to June of 2013 and in March of 2015 and analyzed in order to evaluate the influence of climate, colony of origin, and food supplementation of colonies on the content of total phenolic and flavonoid by chromatographic analysis and antioxidant activity by radical scavenging of 2,2-diphenyl-1-picrylhydrazyl hydrate (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and ferric reducing antioxidant power (FRAP) methods. The Principal Component Analysis (PCA) was carried out with propolis collected in 2013 and two clusters were formed. Propolis produced in the months of March and April showed a higher content of total phenolic compounds (TPC) and antioxidant capacity than those produced in May and June. The results of PCA obtained from samples collected in March of 2013 and 2015 showed two clusters, and propolis collected in 2015 were more bioactive and presented a higher content of TPC. The chromatographic analysis of extracts allowed the identification of phenolic acids p-coumaric, ferulic and caffeic with similar chemical profiles that could be closely related to the botanical origin of propolis. It can be concluded that the season and food supplementation of colonies influenced the chemical composition and the biological activity of samples analysed.

Melipona mondury produces a geopropolis with antioxidant, antibacterial and antiproliferative activities

ABSTRACT Geopropolis is a special type of propolis produced by stingless bees. Several pharmacological properties have been described for different types of geopropolis, but there have been no previous studies of the geopropolis from Melipona mondury. In this study, we investigated the antioxidant, antibacterial, and antiproliferative activities of M. mondury geopropolis, and determined its chemical profile. The antioxidant activity was determined using in vitro ABTS·+, ·DPPH, and β-carotene/linoleic acid co-oxidation methods. The antibacterial activity was determined using a microdilution method with Pseudomonas aeruginosa, Staphylococcus aureus, and methicillin-resistant S. aureus. The antiproliferative effect was determined in tumor cell lines using the Alamar Blue assay. The chemical profile was obtained using UHPLC-MS and UHPLC-MS/MS. The butanolic fraction had the highest concentration of phenolic compounds and more potent antioxidant properties in all assays. This fraction also had bacteriostatic and bactericidal effects against all bacterial strains at low concentrations, especially S. aureus. The hexane fraction had the highest antiproliferative potential, with IC50 values ranging from 24.2 to 46.6 µg/mL in HL-60 (human promyelocytic leukemia cell) and K562 (human chronic myelocytic leukemia cell), respectively. Preliminary chemical analysis indicates the presence of terpenes and gallic acid in the geopropolis. Our results indicate the therapeutic potential of geopropolis from M. mondury against inflammatory, oxidative, infectious, and neoplastic diseases.

Pythium insidiosum: inhibitory effects of propolis and geopropolis on hyphal growth

Abstract Propolis and geopropolis are resinous products of bees showing antimicrobial effects. There is no data concerning their action against Pythium insidiosum - the causative agent of pythiosis, a pyogranulomatous disease of the subcutaneous tissue that affects mostly horses, dogs and humans. Fragments of 15 isolates of P. insidiodum were incubated with propolis and geopropolis extracts and evaluated for up to seven days to detect the minimal fungicidal concentration (MFC). Propolis inhibited three isolates at 1.0 mg mL-1 after 24 h and all other isolates at 3.4 mg mL-1. Geopropolis led to more variable results, exerting predominantly a fungistatic action than a fungicidal one. Propolis was more efficient than geopropolis in inhibiting P. insidiosum since lower concentrations led to no growth after 24 h. This effect may be due to propolis chemical composition, which has more active compounds than geopropolis. Propolis seemed to be a good candidate for in vivo studies, since treatment with conventional antifungal compounds is difficult in most of the cases, requiring extensive surgical debridement.

Effects of seasonal variations and collection methods on the mineral composition of propolis from Apis mellifera Linnaeus Beehives / Efeitos de variações sazonais e métodos de coleta sobre a composição mineral de própolis de colmeias de Apis mellifera Linnaeus

Abstract The effects of seasonal variations and the methods of collection of propolis produced by Africanized honey bees Apis mellifera Linnaeus, 1758, on the composition of constituent minerals such as magnesium (Mg), zinc (Zn), iron (Fe), sodium (Na), calcium (Ca), copper (Cu), and potassium (K) were evaluated. Propolis was harvested from 25 beehives by scraping or by means of propolis collectors (screen, “intelligent” collector propolis [ICP], lateral opening of the super [LOS], and underlay method). During the one-year study, the propolis produced was harvested each month, ground, homogenized, and stored in a freezer at -10 ºC. Seasonal analyses of the mineral composition were carried out by atomic absorption spectrophotometry and the results were evaluated by analysis of variance (ANOVA), followed by Tukey-Kramer’s test to compare the mean values (p<0.05). The results showed that seasonal variations influence the contents of 5 minerals (Mg, Fe, Na, Ca, and Cu), and the propolis harvesting method affects the contents of 4 minerals (Mg, Zn, Fe, and Ca).

Resumo A influência da sazonalidade e de métodos de produção de própolis por abelhas africanizadas Apis mellifera Linnaeus, 1758, sobre a concentração de magnésio (Mg), zinco (Zn), ferro (Fe), sódio (Na), cálcio (Ca), cobre (Cu) e potássio (K) foram avaliados. 25 colmeias foram utilizadas, e a colheita de propolis ocorreu por raspagem ou a partir de coletores (tela, coletor de própolis “inteligente” – CPI, abertura lateral da melgueira – ALM e calço). Durante um ano a própolis foi colhida mensalmente, homogeneizada e armazenada em freezer a -10 ºC. A análise sazonal de minerais foi realizada por espectrofotometria de absorção atômica e os resultados avaliados por análise de variância (ANOVA) seguida do teste de Tukey-Kramer para comparação de médias (p<0,05). Os resultados demostraram que a sazonalidade afetou o conteúdo de cinco minerais (Mg, Fe, Na, Ca e Cu) e os métodos de coleta afetaram o conteúdo de quatro minerais (Mg, Zn, Fe e Ca).

Physicochemical properties and antioxidant capacity of propolis of stingless bees (Meliponinae) and Apisfrom two regions of Tocantins, Brazil / Propriedades físico-químicas e capacidade antioxidante de própolis de abelhas sem ferrão (Meliponinae) e Apis de duas regiões do Tocantins, Brasil

The composition of propolis depends on time, vegetation and the location of the collection area. The objective of this study was to determine the physicochemical characteristics, the concentration of phenol compounds and the antioxidant capacity of propolis of native stingless bees (Meliponinae) and Apis from the State of Tocantins. Extraction with 80% ethanol (v/v) was performed in order to obtain the extracts. Parameters examined were: propolis mass loss by desiccation at 105 ºC, ashes, wax concentration and pH. Furthermore, the propolis antioxidant activity was measured, as well as the total concentration of phenol compounds. The extracts were also analyzed by high performance liquid chromatography. The total concentration of phenol compounds varied between 121.78 and 631.29 (mg GAE g-1). The antioxidant activity expressed by the value of CE50 varied between 29.81 and 845.38 µg mL-1. High performance liquid chromatography analysis allowed us to infer the existence of phenol compounds. The results indicated that the studied propolis samples constitute good sources of natural antioxidants. The variety of phenol compounds identified in this study, and the diverse biological functions reported in literature for these compounds indicated that this stingless bee propolis (Meliponinae) and Apis has a high pharmacological potential.

A composição da própolis depende do tempo, da vegetação e a localização da área de coleta. Este estudo teve como objetivo determinar as características físicas e químicas, o teor de compostos fenólicos e capacidade antioxidante da própolis de abelhas nativas sem ferrão (Meliponinae) e Apis do Estado do Tocantins. Para a obtenção dos extratos empregou-se extração com etanol 80% (v/v). Os parâmetros analisados foram: perda de peso por dessecação a 105 °C, cinzas, Teor de cera e pH. Além dessas foi mensurada a atividade antioxidante e o teor de compostos fenólicos totais. Adicionalmente os extratos de própolis foram também analisados por cromatografia líquida de alta eficiência. O teor de compostos fenólicos totais variou entre 121,78 e 631,29 (mg GAE g-1). A atividade antioxidante expressa pelo valor de CE50 variou entre 29,81 e 845,38 µg mL-1. As análises por cromatografia líquida de alta eficiência permitiram inferir a existência de compostos fenólicos. Os resultados indicaram que as amostras de própolis estudadas constituem boas fontes de antioxidantes naturais. A variedade de compostos fenólicos identificada neste estudo, e as diversas funções biológicas relatadas na literatura para estes compostos, indicaram que a própolis de abelhas sem ferrão (Meliponinae) e Apis tem um grande potencial farmacológico.

Isoflavone formononetin from red propolis acts as a fungicide against Candida sp

Abstract A bioassay-guided fractionation of two samples of Brazilian red propolis (from Igarassu, PE, Brazil, hereinafter propolis 1 and 2) was conducted in order to determine the components responsible for its antimicrobial activity, especially against Candida spp. Samples of both the crude powdered resin and the crude ethanolic extract of propolis from both locations inhibited the growth of all 12 tested Candida strains, with a minimum inhibitory concentration of 256 µg/mL. The hexane, acetate and methanol fractions of propolis 1 also inhibited all strains with minimum inhibitory concentration values ranging from 128 to 512 µg/mL for the six bacteria tested and from 32 to 1024 µg/mL for the yeasts. Similarly, hexane and acetate fractions of propolis sample 2 inhibited all microorganisms tested, with minimum inhibitory concentration values of 512 µg/mL for bacteria and 32 µg/mL for yeasts. The extracts were analyzed by HPLC and their phenolic profile allowed us to identify and quantitate one phenolic acid and seven flavonoids in the crude ethanolic extract. Formononetin and pinocembrin were the major constituents amongst the identified compounds. Formononetin was detected in all extracts and fractions tested, except for the methanolic fraction of sample 2. The isolated isoflavone formononetin inhibited the growth of all the microorganisms tested, with a minimum inhibitory concentration of 200 µg/mL for the six bacteria strains tested and 25 µg/mL for the six yeasts. Formononetin also exhibited fungicidal activity against five of the six yeasts tested. Taken together our results demonstrate that the isoflavone formononetin is implicated in the reported antimicrobial activity of red propolis.

Actividad antibacteriana in vitro de propóleos chilenos sobre Helicobacter pylori / In vitro antibacterial activity of Chilean propolis against Helicobacter pylori

Introduction: Propolis is a natural product derived from beekeeping. It has anesthetic, anti-inflammatory, immune-stimulant and antibacterial properties on grampositive and gramnegative bacteria. However, little is known regarding its activity on Helicobacter pylori. This bacteria colonizes about half of the world’s population and is associated with chronic gastritis, peptic ulcer and gastric cancer. Objective: The aim of this study was to evaluate the inhibitory activity of 22 propolis extracts from nine of the 11 beekeeping Chilean regions on 10 strains of H. pylori isolated from gastric mucosa. Methods: The antibacterial activity of the extracts was determined using the well diffusion method and diffusion disks. Results: 100% of the extracts were active on the tested strains, showing inhibition halos equal to or greater than 15 mm by both methods. Conclusions: our results show an effective anti H. pylori activity of propolis. However, additional microbiological studies are needed before a potential clinical utility of these natural products is warranted.

Introducción: El propóleos es un producto natural derivado de la apicultura que tiene propiedades anestésicas, anti-inflamatorias, inmuno-estimulantes y antibacterianas. Ejerce su acción sobre distintas bacterias grampositivas y gramnegativas. Sin embargo, es muy poco lo que se sabe en relación a su actividad sobre H. pylori, bacteria asociada con gastritis crónica, úlcera gastro-duodenal y cáncer gástrico y que coloniza a alrededor de la mitad de la población mundial. Objetivo: Evaluar la actividad inhibitoria de 22 extractos de propóleos de orígenes botánicos diferentes, provenientes de nueve de las once zonas mielíferas de Chile, en la época de otoño, sobre 10 cepas de H. pylori aisladas de mucosa gástrica. Metodología: La actividad antibacteriana de los extractos se determinó a través del método de difusión en pocillos y de difusión en discos. Resultados: 100% de los extractos fueron activos sobre las cepas ensayadas, observándose halos de inhibición iguales o mayores a 15 mm en ambos métodos. Conclusiones: Los datos obtenidos in vitro en el presente estudio muestran una efectiva actividad anti H. pylori de los propóleos chilenos, siendo necesario estudios microbiológicos y farmacológicos adicionales para avanzar en una posible utilidad clínica de estos productos naturales.

O efeito do uso de fitoterápicos e da própolis nas propriedades físico-químicas, antimicrobiana e biocompatibilidade do MTA / The effect of the use of phytotherapics and propolis in physicochemical properties, antimicrobial and biocompatibility of MTA

O objetivo deste estudo foi avaliar as propriedades físico-químicas, antimicrobianas e biocompatibilidade do MTA branco manipulado com extratos aquoso e/ou em propilenoglicol da Arctium lappa L., Casearia sylvestris Sw. e própolis. Dentre os testes físico-químicos foram avaliados o tempo de presa, escoamento, pH, liberação de íons cálcio e alteração volumétrica. Para verificar o efeito antimicrobiano foram aplicadas as metodologias do contato direto (Enterococcus faecalis e a Cândida albicans) e da descontaminação dentinária, empregando a microscopia confocal de varredura laser para verificar a viabilidade de Enterococcus faecalis. Para a avaliação da biocompatibilidade, 162 ratos Wistar foram utilizados, onde cada animal recebeu dois implantes subcutâneos e um alveolar. Após os períodos experimentais de 15, 30 e 60 dias foram realizadas análises microtomográfica, histológica descritiva e histomorfométrica. Adicionalmente amostras do tecido alveolar foram processadas para dosagem das citocinas TNF-α e IL-10 por meio do ensaio imunoenzimático (ELISA). Os dados obtidos foram analisados estatisticamente com os testes ANOVA e Tukey ou Kruskal-Wallis e Dunn. Os resultados revelaram que a variação do veículo associado ao MTA aumentou significativamente o tempo de presa, no entanto, não houve influência na alteração volumétrica (P>0,05) e na capacidade do cimento em manter o meio alcalino e liberar íons cálcio. Os cimentos manipulados com extratos em propilenoglicol apresentaram maior escoamento (P<0,05). Apenas o extrato da própolis agregou ao MTA efeito contra o Enterococcus faecalis após 24 e 48 horas (descontaminação dentinária e contato direto respectivamente) e contra a Cândida albicans após 10 horas (P<0,05). De acordo com as avaliações histológica e histomorfométrica dos implantes em tecidos subcutâneo e alveolar não foi constatada diferença significativa entre os grupos experimentais quando comparados com o grupo no qual o MTA foi manipulado...

The aim of this study was to evaluate the physicochemical, antimicrobial properties and biocompatibility of white MTA mixed with aqueous or propylene glycol extracts of Arctium lappa L., Casearia sylvestris Sw. and propolis. Among physicochemical tests were evaluated the setting time, flowability, pH, ion calcium release and volumetric change. To verify the antimicrobial effects were applied the methods of direct contact (Enterococcus faecalis and Candida albicans) and dentin decontamination by using the confocal laser scanning microscopy to verify the Enterococcus faecalis viability. To evaluate the biocompatibility were used 162 Wistar rats. Each animal received one alveolar and two subcutaneous implants. After the experimental periods of 15, 30 and 60 days were performed the microtomography, histological description and histomorphometric analyses. Additionally alveolar tissue samples were processed for the measurement of TNF-α e IL-10 cytokines by enzyme-linked immunosorbent assay (ELISA). The data were statistically analyzed by the ANOVA and Tukey or KruskalWallis and Dunns tests. The results revealed that the variation of the vehicle associated to MTA significantly increased its setting time, however did not influence the volumetric change (P>0,05) and the cement's ability to maintain the alkaline medium and ion calcium release. Cements mixed with propylene glycol extracts showed higher flowability (P<0,05). Only propolis extract added to MTA the effect against E. faecalis after 24 and 48 hours (dentin decontamination and direct contact respectively) and against Candida albicans after 10 hours (P<0,05). According to the histological and histomorphometric evaluation of the implants in subcutaneous and alveolar tissue was not observed significant differences between the experimental groups in comparison to the reference group (MTA was mixed with distilled water). The microtomography analysis and expression of TNF-α and IL-10 showed...

O efeito do uso de fitoterápicos e da própolis nas propriedades físico-químicas, antimicrobiana e biocompatibilidade do MTA / The effect of the use of phytotherapics and propolis in physicochemical properties, antimicrobial and biocompatibility of MTA

O objetivo deste estudo foi avaliar as propriedades físico-químicas, antimicrobianas e biocompatibilidade do MTA branco manipulado com extratos aquoso e/ou em propilenoglicol da Arctium lappa L., Casearia sylvestris Sw. e própolis. Dentre os testes físico-químicos foram avaliados o tempo de presa, escoamento, pH, liberação de íons cálcio e alteração volumétrica. Para verificar o efeito antimicrobiano foram aplicadas as metodologias do contato direto (Enterococcus faecalis e a Cândida albicans) e da descontaminação dentinária, empregando a microscopia confocal de varredura laser para verificar a viabilidade de Enterococcus faecalis. Para a avaliação da biocompatibilidade, 162 ratos Wistar foram utilizados, onde cada animal recebeu dois implantes subcutâneos e um alveolar. Após os períodos experimentais de 15, 30 e 60 dias foram realizadas análises microtomográfica, histológica descritiva e histomorfométrica. Adicionalmente amostras do tecido alveolar foram processadas para dosagem das citocinas TNF-α e IL-10 por meio do ensaio imunoenzimático (ELISA). Os dados obtidos foram analisados estatisticamente com os testes ANOVA e Tukey ou Kruskal-Wallis e Dunn. Os resultados revelaram que a variação do veículo associado ao MTA aumentou significativamente o tempo de presa, no entanto, não houve influência na alteração volumétrica (P>0,05) e na capacidade do cimento em manter o meio alcalino e liberar íons cálcio. Os cimentos manipulados com extratos em propilenoglicol apresentaram maior escoamento (P<0,05). Apenas o extrato da própolis agregou ao MTA efeito contra o Enterococcus faecalis após 24 e 48 horas (descontaminação dentinária e contato direto respectivamente) e contra a Cândida albicans após 10 horas (P<0,05). De acordo com as avaliações histológica e histomorfométrica dos implantes em tecidos subcutâneo e alveolar não foi constatada diferença significativa entre os grupos experimentais quando comparados com o grupo no qual o MTA foi manipulado...

The aim of this study was to evaluate the physicochemical, antimicrobial properties and biocompatibility of white MTA mixed with aqueous or propylene glycol extracts of Arctium lappa L., Casearia sylvestris Sw. and propolis. Among physicochemical tests were evaluated the setting time, flowability, pH, ion calcium release and volumetric change. To verify the antimicrobial effects were applied the methods of direct contact (Enterococcus faecalis and Candida albicans) and dentin decontamination by using the confocal laser scanning microscopy to verify the Enterococcus faecalis viability. To evaluate the biocompatibility were used 162 Wistar rats. Each animal received one alveolar and two subcutaneous implants. After the experimental periods of 15, 30 and 60 days were performed the microtomography, histological description and histomorphometric analyses. Additionally alveolar tissue samples were processed for the measurement of TNF-α e IL-10 cytokines by enzyme-linked immunosorbent assay (ELISA). The data were statistically analyzed by the ANOVA and Tukey or KruskalWallis and Dunns tests. The results revealed that the variation of the vehicle associated to MTA significantly increased its setting time, however did not influence the volumetric change (P>0,05) and the cement's ability to maintain the alkaline medium and ion calcium release. Cements mixed with propylene glycol extracts showed higher flowability (P<0,05). Only propolis extract added to MTA the effect against E. faecalis after 24 and 48 hours (dentin decontamination and direct contact respectively) and against Candida albicans after 10 hours (P<0,05). According to the histological and histomorphometric evaluation of the implants in subcutaneous and alveolar tissue was not observed significant differences between the experimental groups in comparison to the reference group (MTA was mixed with distilled water). The microtomography analysis and expression of TNF-α and IL-10 showed...

Chemopreventive effect of Brazilian green propolis on experimental dermal carcinogenesis in murine model / Efecto quimiopreventivo del propóleos verde brasileño sobre la carcinogénesis cutánea en el modelo murino experimental

The aim of this study was to assess the effect of oral administration of Hydroalcoholic Extract of Green Propolis (HEGP) on dermal carcinogenesis in rodent model. For the biological assay, we used 36 mice, assigned into 6 groups (n=6): CTR (treated with 100 mg/kg HEGP and no tumor induction), TUM (treated with water and tumor induction), GP10 (treated with 10 mg/kg HEGP and tumor induction), GP50 (treated with 50 mg/kg HEGP and tumor induction) and GP100 (treated with 100 mg/kg HEGP and tumor induction). Cancer induction was performed in the back of the mice by topical application of DMBA. After 16 weeks, mice were euthanized and their backs were submitted to post-mortem histological analysis. The mean number of lesions developed in TUM (4.14±0.89) was significantly higher than in GP10 (2.05±1.02), GP50 (1.8±1.92) and GP100 (2.5±1.73) (p<0.05). The tumors formed in HEGP-treated groups were histologically more differentiated, but only in PV100 in situ lesions were evidenced. Infiltration of anatomical noble structures was less frequent in HEGP-treated groups (p<0.05). Our data suggest that oral administration of HEGP provided partial inhibition of DMBA-induced dermal carcinogenesis, as well as appeared to modulate the differentiation and infiltrative potential of the carcinomas in rodent model.

El objetivo de este estudio fue evaluar el efecto de la administración oral de extracto hidroalcohólico del propóleos verde (HEGP) sobre la carcinogénesis dérmica en modelo de roedores. Para el ensayo biológico, se utilizaron 36 ratones asignados en 6 grupos (n = 6): CTR (tratado con 100 mg/kg HEGP y sin inducción de tumores), TUM (tratada con agua e inducción de tumores), GP10 (tratado con 10 mg/kg HEGP e inducción de tumores), GP50 (tratado con 50 mg/kg HEGP e inducción de tumores) y GP100 (tratado con 100 mg/kg HEGP e inducción de tumores). La inducción de cáncer se llevó a cabo en la región dorsal de los ratones por aplicación tópica de DMBA. Después de 16 semanas, los ratones fueron sacrificados y sus dorsos fueron sometidos a análisis histológico post-mortem. El número medio de lesiones desarrolladas en TUM (4,14±0,89) fue significativamente mayor que GP10 (2,05±1,02), GP50 (1,8±1,92) y gp100 (2,5±1,73) (p<0,05). Los tumores formados en grupos tratados con HEGP fueron histológicamente más diferenciados, pero sólo en PV100 las lesiones in situ fueron manifiestas. La infiltración de las estructuras anatómicas blanco fue menos frecuente en los grupos tratados con HEGP (p<0,05). Nuestros datos sugieren que la administración oral de HEGP proporciona una inhibición parcial de la carcinogénesis dérmica inducida por DMBA, así como pareció modular la diferenciación y potencial infiltrante de los carcinomas en el modelo animal.

Chemical and botanical characterization of Chilean propolis and biological activity on cariogenic bacteria Streptococcus mutans and Streptococcus sobrinus

Propolis is a non-toxic natural substance with multiple pharmacological properties including anticancer, antioxidant, fungicidal, antibacterial, antiviral, and anti-inflammatory among others. The aim of this study was to determine the chemical and botanical characterization of Chilean propolis samples and to evaluate their biological activity against the cariogenic bacteria Streptococcus mutans and Streptococcus sobrinus. Twenty propolis samples were obtained from beekeeping producers from the central and southern regions of Chile. The botanical profile was determined by palynological analysis. Total phenolic contents were determined using colorimetric assays. Reverse phase HPLC and HPLC-MS were used to determine the chemical composition. The minimum inhibitory concentration (MIC) was determined on S. mutans and S. sobrinus. All propolis samples were dominated by structures from native plant species. The characterization by HPLC/MS, evidenced the presence of quercetin, myricetin, kaempferol, rutine, pinocembrin, coumaric acid, caffeic acid and caffeic acid phenethyl ester, that have already been described in these propolis with conventional HPLC. Although all propolis samples inhibited the mutans streptococci growth, it was observed a wide spectrum of action (MIC 0.90 to 8.22 µgmL-1). Given that results it becomes increasingly evident the need of standardization procedures, where we combine both the determination of botanical and the chemical characterization of the extracts. Research conducted to date, describes a promising effectiveness of propolis in the prevention of caries and other diseases of the oral cavity, making it necessary to develop studies to identify and understand the therapeutic targets or mechanisms of molecular action of the various compounds present on them.

Estudo comparativo do uso tópico de própolis verde e vermelha na reparação de feridas em ratos / Comparative study of topical green and red propolis in the repair of wounds induced in rats

OBJETIVO: Avaliar a ação cicatrizante de própolis verde e vermelha, correlacionando ao teor de flavonoides MÉTODOS: Foi realizada a quantificação de flavonoides totais dos extratos etanólicos de própolis verde e vermelha para posterior incorporação em pomada base a 20%. Utilizou-se 20 ratos Wistar distribuídos em quatro grupos: solução salina a 0,9% (S), pomada base (B), pomada própolis verde (G), pomada própolis vermelha (R), todos submetidos à confecção de lesões excisivas na região mediana do dorso. Os ratos foram tratados diariamente durante 15 dias. Neste período foram observados peso; temperatura corporal; diâmetro das feridas. Para análise histológica, amostras das feridas foram coletadas. Ao fim do experimento foram realizadas coleta sanguínea e remoção do rim e fígado para análises bioquímica e histológica. RESULTADOS: Os teores de flavonoides totais das própolis verde (4,50 %) e vermelha (5,92 %) foram elevados (>2 %), mas, embora a segunda apresente teor maior que a primeira, a evolução da própolis verde, macro e histologicamente, foi melhor na reparação das feridas. Não foram observadas nefro ou hepatotoxicidade, resultado corroborado pelos ensaios bioquímicos (TGP e albumina). A própolis influiu na redução de colesterol total, triglicerídeos e glicemia. CONCLUSÃO: Não foi possível correlacionar o teor de flavonoides totais com a ação cicatrizante da própolis. O dado revela a necessidade da elucidação dos flavonoides encontrados em cada classe de própolis para desvendar qual (ou quais) flavonoide(s) seria(m) representativo(s) no processo cicatricial.

OBJECTIVE: To evaluate the healing action of green and red propolis, correlating it with the content of flavonoids. METHODS: We performed quantification of total flavonoids of green and red propolis ethanol extracts for subsequent incorporation in ointment base to 20%. We used 20 Wistar rats divided into four groups: 0.9% saline (S), ointment base (B), green propolis ointment (G) and red propolis ointment (R). All animals were submitted to excisional lesions in the midian back region. The rats were treated daily for 15 days. During this period we observed weight, body temperature and diameters of the wounds. For histological analysis, samples were collected from wounds. At the end of the experiment we performed blood collection and removal of the kidney and liver for biochemical and histological analyzes. RESULTS: The levels of total flavonoids of green (4.50%) and red (5.92%) propolis were high (> 2%), but, while the latter showed a content larger than the former, the evolution of green propolis was better in the repair of wounds, both macroscopically and histologically. There were no nephrotoxicity or hepatotoxicity, a result confirmed by biochemical tests (ALT and albumin). Propolis influenced the reduction of total cholesterol, triglycerides and glucose. CONCLUSION: There was no correlation between total flavonoid contents and the healing action of propolis. This reveals the need for elucidation of the flavonoids found in each class of propolis to unravel which one(s) would be important for the healing process.