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

Increased Expression of RUNX2 and ALP Resulting from a Combination of Propolis Extract and Bovine Bone Graft in Socket Preservation Material

Abstract The aim of this study was to determine the inductive effect of a combination of propolis and BBG extract on RUNX2 and ALP expression in the tooth extraction sockets of Cavia cobaya. Fifty- six Cavia cobaya were divided into four groups: polyethylene glycol (PEG), propolis extract + PEG, BBG + PEG, and propolis extract + BBG + PEG. The lower left incisor was extracted, and the socket subsequently filled with material according to the specific group of which the subject was a member. The subjects were sacrificed on the 14th and 30th days. Immunohistochemical staining was carried out under a light microscope at 400x magnification. Statistical analysis was then carried out by means of One-Way ANOVA and Tukey HSD tests. The mean number of RUNX2 and ALP expressions in each group was significantly different. The highest number of RUNX2 and ALP expressions occurred in the propolis + BBG + PEG group on the 30th day, while the lowest expressions were observed in the control group on the 14th day. A combination of propolis and BBG extract at a concentration of 2% of active substance effectively increases the expression of RUNX2 and ALP in preserving the tooth extraction sockets of Cavia cobaya.

Application of propolis in agriculture

This work carried out a literary review of the different aspects related to the use of propolis in the management of plant crops and their advances in agriculture. Propolis is a product widely known for its therapeutic capacity. Due to its antimicrobial and antioxidant properties, among other biological activities, its use has been studied in agriculture. Studies they show that the use of propolis via ethanolic propolis extract (EEP) provides beneficial effects on crops. These benefits are associated with the control of phytopathogens, postharvest preservation and conservation of fruits and vegetables and promotion of plant growth. There is evidence that benefits are the result of the propolis chemical composition that is especially rich in phenolic compounds and nutrients. In addition, the typical waxy property of propolis results in an efficient biofilm in plant tissues. There are still scarce reports showing that the use of EEP in crop management controls the presence of insects and reduces water stress in plants. Generally, studies are mainly focused on in vivo and greenhouse evaluations, requiring further research to elucidate the full potential of the use of propolis in crop management.

Antimicrobial activity of propolis extract fractions against Staphylococcus spp. isolated from goat mastitis / Atividade antimicrobiana de frações do extrato de própolis frente à Staphylococcus spp. isolados de mastite caprina

The indiscriminate use of antibiotics in the treatment of caprine mastitis causes the appearance of resistant microorganisms, besides leaving residues in milk, putting at risk to human health. In this way, propolis is an alternative in the treatment of diseases because it has antimicrobial activity, mainly because of the presence of flavonoids in its composition. The aim of this study was to evaluate the antimicrobial potential of propolis to Staphylococcus spp. Isolated from cases of goat mastitis and qualify the crude ethanoic extract by high performance liquid chromatography (HPLC). In this study, the minimum bactericidal concentration values of propolis extracts in ethanol, ethyl acetate and hexane showed that the best concentrations capable of promoting the highest mortality of the isolates of Staphylococcus spp. from mastitis in goats, were 6250, 3125 and 1562.5µg/mL, respectively. By the microplate adherence test, it was found that 20.78% isolates were not able to form biofilm, 14.70% were classified as moderate and 64.70% were weak and none as a strong biofilm producer. Propolis in its different diluents was able to affect the formation of biofilm and showed a pronounced marked antimicrobial activity against Staphylococcus spp. strains and may be indicated for use in in vivo studies.(AU)

O uso indiscriminado de antibióticos no tratamento de mastite caprina leva ao desenvolvimento de micro-organismos resistentes que poderão estar presentes em alimentos, colocando em risco a saúde humana. Dessa forma, a própolis surge como uma alternativa para o tratamento de doenças por possuir uma ação antimicrobiana, principalmente pela presença de flavonoides em sua composição. O objetivo desse estudo foi avaliar o potencial antimicrobiano da própolis frente à Staphylococcus spp. isolados de casos de mastite caprina e qualificar o extrato etanoico bruto por cromatografia líquida de alta eficiência (CLAE-DAD). Neste estudo, os valores de concentração bactericida mínima (CBM) dos extratos de própolis em álcool etílico, acetato de etila e hexano nos isolados foram de 6250, 3125 e 1562,5µg/mL, respectivamente. Pelo teste de aderência à microplacas, observou-se que 20,78% dos microorganismos, não foram capazes de formar biofilme, 14,70% foram classificados como moderados, 64,70% em fracos e nenhum como forte produtor de biofilme. A própolis em seus diferentes diluentes foi capaz de afetar a formação de biofilme e apresentou significativa atividade antimicrobiana frente a cepas de Staphylococcus spp., podendo ser indicada para utilização em estudos "in vivo".(AU)

Preparation technology of propolis extract / 中草药

Objective: To establish the preparation technology of propolis extract. Methods: With the contents of total flavonoids, chrysin, pinocembrin, and galangin and extract yield as evaluation indexes, the extraction methods, extraction solvent, extraction times, extraction time, and amount of solvent extraction were optimized with single factor test and orthogonal design to investigate the effect on the preparation technology of propolis extract. Results: The optimal extraction technology was that propolis was refrigerated, grinded, soaked with 95% five times the amount of ethanol, shaked, then extracted for 4 h, filtrated, concentrated, and dried after ethanol recovery. Conclusion: The optimial extracting technology of propolis is stable and feasible, and suitable for further application.

Antioxidant effect of aqueous extract of propolis on hepatotoxicity induced by octylphenol in male rats / Efecto antioxidante del extracto acuoso de propóleos sobre la hepatotoxicidad inducida por el octilfenol en ratas macho

4-tertiary-octylphenol (4-tert-OP) is an alkylphenol that affects human health by stimulating free radical production. Aqueous propolis extract is a natural product rich in favonoids that have antioxidant activity. This study was designed to investigate the ability of aqueous propolis extract to reduce the hepatotoxicity induced by 4-tert-OP in male rats. Animals were assigned to 5 groups and treated for 6 weeks. Group 1: control; group 2: 100 mg 4-tert-OP/kg b.wt./day; group 3: 100 mg aqueous propolis extract /kg b.wt./day; group 4: 100mg 4-tert-OP/kg b.wt./day plus 100 mg aqueous propolis extract /kg b.wt./day; group 5: 100 mg 4-tert-OP/kg b.wt./day for 6 weeks followed by 100 mg aqueous propolis extract /kg b.wt./day for 6 weeks. Group 4-tert-OP signifcantly elevated AST, ALT, ALP, GGT, bilirubin, creatinine, urea, total lipids, total cholesterol, triglycerides, LDL-C and MDA with a signifcant decrease in total proteins, albumin, globulin, HDL-C, total antioxidant capacity, SOD, CAT and GST compared to control group. Administration of aqueous propolis extract either alone or combined with 4-tert-OP ameliorated the hepatotoxicity induced by 4-tert-OP. DNA fragmentation supported the deleterious effect of 4-tert-OP and the ameliorative effect of propolis on liver cellular proteins and enzymes. Histopathological fndings revealed the hepatotoxicity induced by 4-tert-OP and the protective effect of aqueous propolis extract. In conclusion, aqueous propolis extract could reduce the damage and toxicity effects on liver cells induced by 4-tert-OP.

El 4-terc-octilfenol (4-terc-OP) es un alquilfenol que afecta a la salud humana mediante la estimulación de la producción de radicales libres. El extracto acuoso de propóleos es un producto natural rico en favonoides que tienen actividad antioxidante. Este estudio fue diseñado para investigar la capacidad del extracto de propóleos de reducir la hepatotoxicidad inducida por el 4-terc-OP en ratas macho. Los animales fueron asignados a 5 grupos y tratados durante 6 semanas. Grupo 1: control; grupo 2: 100 mg de 4-terc-OP/kg/día; grupo 3: 100 mg de extracto de propóleos/kg/día; grupo 4: 100 mg de 4-terc-OP/ kg/día más 100 mg de extracto de propóleos/kg/día, grupo 5: 100 mg de 4-terc-OP/kg/día durante 6 semanas, seguidos de 100 mg de extracto de propóleos/kg/día durante 6 semanas. El grupo 4-terc-OP mostró niveles signifcativamente elevados de AST, ALT, ALP, GGT, bilirrubina, creatinina, urea, lípidos totales, colesterol total, triglicéridos, LDL-C y MDA, con una disminución signi-fcativa de proteínas totales, albúmina, globulina, HDL-C, la capacidad antioxidante total, SOD, CAT y GST, en comparación con el grupo control. La administración de extracto de propóleos, ya sea solo o combinado con 4-terc-OP redujo la hepatotoxicidad inducida por 4-terc-OP. Los estudios de fragmentación del ADN apoyan el efecto deletéreo observado por el tratamiento con 4-terc-OP y el efecto protector del extracto de propóleos, sobre las proteínas y las enzimas celulares hepáticas. Los resultados histopatológicos revelaron la hepatotoxicidad por 4-terc-OP y efecto protector inducido por el extracto de propóleos. En conclusión, el extracto de propóleos podría reducir el daño hepático y los efectos celulares de toxicidad en las células del hígado inducidos por 4-terc-OP.

Atividade antiviral do extrato de própolis contra o calicivírus felino, adenovírus canino 2 e vírus da diarréia viral bovina / Antiviral activity of propolis extracts against feline calicivirus, canine adenovirus 2, and bovine viral diarrhea virus

Dentre as propriedades biológicas da própolis, a atividade antimicrobiana tem merecido destacada atenção. Neste artigo, descreve-se a atividade antiviral de dois extratos etanólicos de própolis (EP1 e EP2) frente aos vírus: calicivírus felino (FCV), adenovírus canino tipo 2 (CAV-2) e vírus da diarréia viral bovina (BVDV). Um dos extratos (EP1) foi obtido por extração etanólica de própolis obtida da região central do Estado do Rio Grande do Sul e o segundo (EP2), obtido comercialmente de uma empresa de Minas Gerais. A análise dos extratos de própolis através da cromatografia líquida de alta eficiência (CLAE) identificou a presença de flavonóides como: rutina, quercetina e ácido gálico. A atividade antiviral bem como a citotoxicidade dos extratos aos cultivos celulares foram avaliadas através do MTT [3- (4,5 dimetiltiazol-2yl)-2-5-difenil-2H tetrazolato de bromo]. Ambos os extratos evidenciaram atividade antiviral frente ao BVDV e CAV-2 quando acrescidos ao cultivo celular anteriormente à inoculação viral. Os extratos foram menos efetivos contra o FCV em comparação aos resultados obtidos com os outros vírus, e a atividade antiviral neste caso foi observada apenas quando a própolis estava presente após a inoculação viral. O extrato obtido no laboratório (EP1) apresentou valores mais altos de índice de seletividade (IS=CC50/ CE50), quando comparado à outra amostra (EP2). Em resumo, a própolis apresentou atividade antiviral frente a três diferentes vírus, o que a torna alvo para o desenvolvimento de novos compostos naturais com atividade antiviral.

Propolis is a resinous substance produced by bees for which several biological activities have been attributed. In this article, the antiviral activity of two propolis extracts was tested against bovine viral diarrhea virus (BVDV), canine adenovirus type 2 (CAV-2), and feline calicivirus (FCV). One of the extracts was obtained by ethanolic extraction of propolis from the Santa Maria (RS) region, while the other was bought from a Minas Gerais industry. The high efficiency liquid cromatography (HPLC) analysis detected the presence of some flavonoids like rutin, quercetin, and gallic acid. The MTT test was applied in order to detect the citotoxicity and also the antiviral activity. Both extracts showed antiviral activity against BVDV and CAV-2 when incubated with the cell cultures before viral inoculation. The extracts were less effective against FCV comparing to the results for the other viruses and, the antiviral activity was observed only when the própolis was present after virus inoculation The extract obtained in the lab showed the highest selectivity index (SI= CC50/ EC50). Thus, propolis showed antiviral activity against three different viruses, making it a target for the development of new natural compounds with antiviral activity.