Propolis is a traditional value-added product from beekeeping, and has been widely used in medicine and the chemical industry because of its extensive biological activities. In this dissertation some pharmacological properties of propolis extracted by ethanol (EEP) and water (WSP) were studied by means of modem pharmacological methods. The results of the experiments show the following: 1. Both EEP and WSD led to decreased levels of fasting blood glucose (FBG), glycosylated haemoglobin (HBA1c), Fructosamine (FRU), blood urea nitrogen (BUN), uric acid (UA), malonaldehyde (MDA), nitric oxide (NO), nitric oxide synthetase (NOS), total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and very-low density lipoprotein cholesterol (VLDL-C) in the serum of fasting rats; and, to increased serum levels of high-density lipoprotein cholesterol (HDL-C) and superoxide dismutase (SOD) in rats with diabetes mellitus. This suggests that propolis can control blood glucose and modulate the metabolism of glucose, blood lipid and protein, leading to decreased outputs of lipid peroxidation and scavenging free radicals in rats with diabetes mellitus. 2. Both EEP and WSD showed inhibitory effects on swelling induced by Freund's complete adjuvant and decreased the degree of local inflammatory responses; significantly inhibited the increase of interleukin-6 (lL-6) in inflamed tissues, but had no significant effect on levels of interleukin-2 (IL-2) and interferon-y (IFN-y). The results are consistent with the interpretation that EEP and WSD may exert these effects by inhibiting the activation and differentiation of mononuclear macrophages. 3. Both EEP and WSD exhibited significant anti-inflammatory effects in animal models with respect to thoracic capillary vessel leakage in mice, Carrageenan-induced oedema, Carrageen-aninduced pleurisy and acute lung damage in rats. The mechanisms for the anti-inflammatory effects probably involve decreasing prostaglandin and NO levels. 4. Both EEP and WSD had an inhibitory effect on the level of TG, TC, LDL-C, glutamic-pyruvic transaminase (GPT) and glutamic-oxalacetic transaminase (GOT) in serum, and TC, TG and MDA in liver of hyperlipidemic SD rats; but were without effects on HDL-C, MDA, SOD and NO in serum. EEP also reduced body weight, liver weight and liver index, but WSD did not reduce those indexes. The results showed that the two extracts contribute to the improvement of lipid metabolism in hyperlipidemic rats and provide them with the required anti-oxidative activity. 5. The 80% ethanol extracts of propolis had the highest flavonoid content. The flavone content of water extracts of propolis were obtained with a natural surface-active agent, under ultrasonic perturbation at 80°C, for 12 h and was 6.7 times greater than that of propolis extracted with water at room temperature. Compared to EEP, WSD has the same or a greater anti-tumor or anti-inflammation effect at the same dosage, and shows considerable pharmacological potential especially because of its low side effects and ease of preparation.

This study was conducted to investigate a method for stimulating increased propolis production in Cape honeybee hives. The study took place near George situated in the Southern Cape region of South Africa. Standard Langstroth hives were used (n = 12) containing honeybee colonies of equal strength from the species Apis mellifera capensis. Propolis production in colonies equipped with commercial propolis traps (n = 6) was compared to propolis production in control colonies (n = 6). The strength of the colonies was determined by calculating the area of worker brood cells and the number of frames containing bees. For a ninemonth period propolis production, honey production, brood area, resin percentage, frames containing bees and income generated were measured on a monthly basis with the removal of the traps. There were no significant difference (p > 0.5) in honey production (24.66 ± 1.19 kg and 26.53 ± 1.31 kg), worker brood area (1967.3 ± 258.61 cm2 and 1085.4 ± 312.99 cm2) and frames containing bees (10 ± 1.13 frames and 9.8 ± 1.2 frames) between the hives with traps and hives without traps. However, propolis production was significantly influenced (p < 0.01) by the propolis traps, with the hives containing traps producing 361.87 ± 8.78g propolis compared to 38.2 ± 2.17g propolis in the control group. Propolis production in both control (28 g) and treatment groups (260.2 g) was significantly influenced by honey flow (p = 0.027). From December to March the production of honey was 22.12 (control) and 24.25 kg (treatment). A positive correlation was also found between frames containing bees and worker brood area (R = 0.73). Hives containing propolis traps were more profitable than the control group when honey and propolis income were pooled (467.17 ± 19.47 Rand vs. 353.8 ± 17.03 Rand). Therefore, the increased propolis production significantly improved profitability of the hive without affecting overall hive productivity.

O presente trabalho teve por objetivo estudar a atividade antifúngica e antibacteriana de diferentes tipos de própolis, tanto de abelhas africanizadas (verde) quanto meliponíneos. A concentração inibitória mínima, sendo a menor concentração de um agente inibidor em que não se observa o crescimento macroscópico do microorganismo, foi determinada para todas as amostras de própolis. Para análise do perfil químico das amostras foi utilizada como metodologia (CLAE), visando determinar as classes de substâncias presentes nas amostras de própolis. Em todos os microorganismos testados as própolis das abelhas Tetragonisca angustula, Plebeia droryana, Scaptotrigona bipunctata, Friesiomelitta varia e Nannotrigona testaceicornis não apresentaram atividade antifúngica nem antibacteriana. Nenhuma das amostras de própolis testadas apresentou atividade antifúngica frente ao Aspergillus nidulans. A linhagem do fungo Trichophyton rubrum mostrou-se sensível a alguns tipos de própolis sendo que a própolis verde foi mais eficaz. Alguns tipos de própolis apresentaram atividade antibacteriana, indicando a importância dessas resinas na terapêutica humana e veterinária. / Our objective was to examine the antifungal and antibacterial activities of several types of propolis, including Africanized honey bee green propolis and propolis produced by meliponini bees. The minimal inhibitory concentration, considered to be the lowest concentration of an inhibitory agent in which there is no macroscopic growth of the microorganism, was determined for all the propolis samples. CLAE methodology was used for the analysis of the chemical profiles of the samples, in order to determine the classes of substances in the propolis samples. Propolis from the bees Tetragonisca angustula, Plebeia droryana, Scaptotrigona bipunctata, Friesiomelita varia and Nannotrigona testaceicornis presented no antifungal or antibacterial activity. None of the propolis samples presented antifungal activity against Aspergillus nidulans. The fungus Trichophyton rubrum was sensitive to several types of propolis, green propolis being the most efficient against this species. Various types of propolis also presented antibacterial activity, demonstrating the potential importance of these resins for human and veterinary medicine.

O presente trabalho teve por objetivo estudar a atividade antifúngica e antibacteriana de diferentes tipos de própolis, tanto de abelhas africanizadas (verde) quanto meliponíneos. A concentração inibitória mínima, sendo a menor concentração de um agente inibidor em que não se observa o crescimento macroscópico do microorganismo, foi determinada para todas as amostras de própolis. Para análise do perfil químico das amostras foi utilizada como metodologia (CLAE), visando determinar as classes de substâncias presentes nas amostras de própolis. Em todos os microorganismos testados as própolis das abelhas Tetragonisca angustula, Plebeia droryana, Scaptotrigona bipunctata, Friesiomelitta varia e Nannotrigona testaceicornis não apresentaram atividade antifúngica nem antibacteriana. Nenhuma das amostras de própolis testadas apresentou atividade antifúngica frente ao Aspergillus nidulans. A linhagem do fungo Trichophyton rubrum mostrou-se sensível a alguns tipos de própolis sendo que a própolis verde foi mais eficaz. Alguns tipos de própolis apresentaram atividade antibacteriana, indicando a importância dessas resinas na terapêutica humana e veterinária. / Our objective was to examine the antifungal and antibacterial activities of several types of propolis, including Africanized honey bee green propolis and propolis produced by meliponini bees. The minimal inhibitory concentration, considered to be the lowest concentration of an inhibitory agent in which there is no macroscopic growth of the microorganism, was determined for all the propolis samples. CLAE methodology was used for the analysis of the chemical profiles of the samples, in order to determine the classes of substances in the propolis samples. Propolis from the bees Tetragonisca angustula, Plebeia droryana, Scaptotrigona bipunctata, Friesiomelita varia and Nannotrigona testaceicornis presented no antifungal or antibacterial activity. None of the propolis samples presented antifungal activity against Aspergillus nidulans. The fungus Trichophyton rubrum was sensitive to several types of propolis, green propolis being the most efficient against this species. Various types of propolis also presented antibacterial activity, demonstrating the potential importance of these resins for human and veterinary medicine.

Berlin, Freie Univ., Diss., 2003. / Dateiformat: zip, Dateien im PDF-Format. Computerdatei im Fernzugriff.

Berlin, Freie Univ., Diss., 2003. / Dateiformat: zip, Dateien im PDF-Format. Computerdatei im Fernzugriff.

Berlin, Freie Universiẗat, Diss., 2003. / Dateiformat: zip, Dateien im PDF-Format.

A dissertation submitted to the Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, in fulfilment of the requirements for the degree of Master of Pharmacy June 2019 / Propolis is a resinous substance produced by the Apis mellifera bee. The compounds predominantly found in South African propolis are the flavonoids pinocembrin, galangin, and chrysin. The aim of this study was to obtain an understanding of the antimicrobial activity of these compounds, both singularly and in combination, and to investigate the role of interactions between the compounds. To observe the effects of the compounds against planktonic micro-organisms, the minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) assays were undertaken. Combinations were tested at equal ratios using the MIC assay, the results of which were interpreted using the fractional inhibitory concentration (FIC) index. Varied ratio combinations using the MIC method were also undertaken and presented in isobolograms. Results from six bacterial strains and three yeast strains demonstrated that the singular compounds displayed moderate to weak activity, and that the activity of pinocembrin, galangin, and chrysin was improved when used in combination. When tested at 1:1 ratio, synergy occurred in six out of the 27 combinations (22%) whilst nine combinations (30%) were additive, and the remaining 12 combinations (44%) were non-interactive. No combination showed antagonism. The 1:1 combination of galangin and chrysin appeared to be the most effective, as it was synergistic against three of the nine micro-organisms (33%) tested, and additive against another three. Pinocembrin, galangin, and chrysin were then tested at varying ratio combinations against three micro-organisms, which produced nine isobolograms and a total of 81 ratios. From these 81 ratios, 32 ratios (40%) showed synergy, while 31 ratios (38%) were additive and the remaining 18 ratios (22%) were non-interactive. The combination of galangin and chrysin against C. tropicalis displayed the best activity, as all nine ratios of these two compounds in combination, showed synergy. The triple combination which used pinocembrin, galangin, and chrysin at a 1:1:1 ratio against nine pathogens, produced synergy against six of the nine micro-organisms (67%) tested and additivity against the remaining three (33%) micro-organisms. Selected combinations 4 possessed bactericidal activity, although the compounds on their own demonstrated no bactericidal activity at the concentrations tested. The most pronounced bactericidal activity was observed from the combination of galangin with chrysin against Candida tropicalis, with an MBC value of 0.16 mg/ml. Anti-quorum sensing (QS) testing was undertaken using Chromobacterium violaceum as a monitor strain. The broth macrodilution method was used. In addition, an anti-QS broth microdilution method was tested for the first time in this study. Results from the broth macrodilution method showed that pinocembrin, galangin and chrysin, and their combinations, were capable of inhibiting violacein production from C. violaceum. When tested singularly, pinocembrin, galangin and chrysin all showed minimum quorum sensing inhibitory concentration (MQSIC) values of 0.31 mg/ml or less, and MBC values of 2.5 mg/ml or less. When tested together, the combinations of pinocembrin, galangin and chrysin showed MQSIC values of 0.16 mg/ml or less, and MBC values of 1.25 mg/ml or less. The combination of pinocembrin and galangin was the most promising, with a MQSIC value of 0.08 mg/ml and an MBC values of 0.31 mg/ml. Three out of four combinations (75%) demonstrated synergy through inhibiting QS, while the combination of pinocembrin and chrysin was additive. Results from the broth microdilution method corresponded with those of the macrodilution method qualitatively, although the quantitative results differed. To assess the effects of pinocembrin, galangin and chrysin on biofilm prevention and disruption, the crystal violet (CV) assay was used. No clear pattern on antibiofilm activity was observed, although selected compounds and combinations were shown to be effective in inhibiting biofilms. Good biofilm inhibitory activity was noted for 18 out of 45 (40%) of the studies when the compounds were tested independently against the three micro-organisms at various timeframes. The best activity from a single compound was observed for chrysin against Escherichia coli at 24 hr (79.73% inhibition). When the compounds were tested in combination at a 1:1 ratio, only nine out of 60 (15%) combinations tested showed good inhibitory activity. However, the triple combination showed the best inhibitory activity of 100% when tested against E. coli at 24 hrs. The toxicity of pinocembrin, galangin and chrysin was screened using the brine shrimp lethality assay (BSLA) after an exposure period of 24 hr. None of the compounds or combinations displayed 5 toxicity. When tested independently, the highest percentage mortality was noted from galangin, at 23.53%. When the compounds were tested in combination, the highest percentage mortality was observed for the combination of galangin and chrysin, at 6.17%. Three of the four combinations showed reduced toxicity. This study has clearly demonstrated that the combination of the compounds pinocembrin, galangin and chrysin found in South African propolis synergistically enhances the antimicrobial activity of South African propolis on a number of levels whilst simultaneously further reducing the toxicity of the compounds. Furthermore, this study provides a convincing example of the need to examine compound interactions and to not always follow the reductionist approach of searching for a single active compound in natural product research. / E.K. 2019

Abstract Propolis has been used by man for millennia for its antimicrobial and pharmaceutical properties. However, its use as an agricultural antimicrobial agent has only recently been assessed. This study assessed the use of an ethanolic extract of propolis (EEP) for the control of avocado fruit fungal pathogens. Qualitative analyses of EEP indicated flavonoids as the main antimicrobial constituents. Quantitative analyses detected 16.35 mg ml-1 total flavonoids and 3.28 mg ml-1 total phenolics. The Minimum Inhibitory Concentration (MIC) of EEP was determined as 5 mg ml-1 against Colletotrichum gloeosporioides, Pestalotiopsis guipinii, a complex of Colletotrichum gloeosporioides and Pseudocercospora sp. (CgP complex), Verticillium sp., Fusarium sp. and Monilia sp., isolated from avocado fruits, using the agar dilution method, at a concentration gradient from 1 to 10 mg ml-1. Electron micrographs of Pestalotiopsis guipinii, Colletotrichum sp. and Colletotrichum gloeosporioides/Pseudocercospora sp. (CgP complex) incubated on agar media containing EEP clearly indicated signs of cell wall damage with large pores within the hyphae. Conidial germination of Colletortichum sp. and P. guipinii was inhibited by 98.95 % and 40.41 % respectively by EEP. Trees infected with Colletotrichum sp., P. guipinii or CgP complex conidia were incubated within greenhouse conditions and treated with 5 mg ml-1 EEP at weekly intervals from once every week to once every six weeks. Disease indices from experimental and control trees were similar but noticeable control of CgP disease symptoms was observed from treatment with EEP. ‘Fuerte’ avocado trees were treated with copper hydroxide, borehole water or 5 mg ml-1 EEP during the 2006-2007 growing season. All fruit were similar after harvesting with respect to pre-harvest disease. The occurrence of post-harvest diseases was analysed after simulations of import and export markets. EEP treated fruit were similar to bore-hole treated fruits (control). Similar results were observed after trials to assess the use of EEP as a post-harvest dip. The stem-end of prematurely harvested ‘Fuerte’ fruit were dipped into 5 mg ml-1 EEP, and incubated until ripe, to assess control of stem-end rot (SER). EEP reduced the occurrence of SER by 30 %. EEP was further assessed to inhibit infection or disease spread by Colletotrichum conidia. Fruits were inoculated with Colletotrichum conidia and either treated with 5 mg ml-1 or 10 mg ml-1 EEP either after or before infection. The fruits were incubated until ripe. Both treatments reduced the occurrence of disease (P < 0.001). In conclusion, EEP did not efficiently control disease in the field, but showed high potential as a future fungicide for avocado fruit. Optimisation of EEP includes higher concentrations, the addition of stickers, and/or more frequent spraying of trees.

Resumo: O consumo de produtos apícolas tem atraído o interesse da população devido à prática de medicinas naturais. As abelhas coletam substâncias e fragmentos de origem vegetal para produzirem a própolis, que é utilizada para selar e proteger a colméia contra invasões de microrganismos patogênicos. O homem utiliza a própolis desde a antiguidade e para fins diversos. Por meio de extensivos estudos, hoje se sabe que existem tipos específicos de própolis, que variam entre as regiões geográficas e a fonte botânica e, portanto, apresentam composições químicas também específicas. A própolis do tipo verde, que ganhou, recentemente, destaque internacional pela sua alta e variada constituição de flavonóides, tem como fonte botânica a planta Baccharis dracunculifolia (Asteraceae), popularmente conhecida como alecrim-do-campo. Atualmente, extratos etanólicos de produtos naturais, como da própolis verde, vêm se destacando pelas suas propriedades terapêuticas. Neste trabalho foi avaliado o potencial citotóxico, genotóxico e mutagênico, mas, principalmente, foi verificada a possível atividade anticitotóxica, antigenotóxica e antimutagênica destes extratos etanólicos, sobre sistemas-teste de Allium cepa (cebola) e células de hepatoma de rato (HTC), mantidas em cultura. A verificação do índice mitótico, de células meristemáticas de A. cepa, não serviu de parâmetro para avaliar citotoxicidade e anticitotoxicidade. As análises da freqüência de aberrações cromossômicas e da freqüência de micronúcleos (MN) revelaram que, ambos os extratos, não foram genotóxicos e nem mutagênicos para as células meristemáticas de A. cepa. Também não houve indução significativa de MN para as células F1 de cebola e para as células HTC mantidas em cultura. O ensaio do cometa, realizado com as células HTC, indicou a ausência de ação genotóxica para os extratos testados... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Consumption of apicultural products has attracted the interest of the population due to the practice of natural medicine. Bees collect substances and fragments of vegetal origin to produce propolis, which is used to seal and protect the beehive against pathogenic microorganisms. Human beings use the propolis since the antiquity and for several purposes. By extensive studies, nowadays it is known that there are specific types of propolis, which vary between geographic regions and botanical sources and, therefore, present specific chemical compositions. The Brazilian green propolis, which has recently received international prominence, has as botanical source the plant Braccharis dracunculifolia (Asteraceae), popularly known as alecrim-do-campo. Currently, ethanolic extracts of natural products, eg made with green propolis, have been highlighted for their therapeutical properties. In the present work, the cytotoxic, genotoxic and mutagenic potentials were evaluated, but, mainly, it was verified the possible anticytotoxic, antigenotoxic and antimutagenic activities of these ethanolic extracts in the Allium cepa (onion) test system and in HTC (hepatoma tissue culture) cells maintained in culture. The verification of the mitotic index, of meristematics cells of A. cepa, did not serve as parameter to evaluate cytotoxicity and anticytotoxicity. The analyses of chromosomal aberrations (AC) and micronuclei (MN) frequencies revealed that, both extracts, were not genotoxic nor mutagenic for meristematics cells of A. cepa. Also, there was no significant induction of MN in onion F1 cells and HTC cells. The comet assay, carried out with HTC cells, indicated absence of genotoxic action for the extracts tested in this work. The pre-treatment carried out with cells of A. cepa showed that the extracts were capable to reduce the induction of cytogenetics damages promoted by Methyl Methane Sulfonate (MMS)... (Complete abstract click electronic access below) / Orientador: Maria Aparecida Marin-Morales / Coorientador: Osmar Malaspina / Banca: Carmem Silvia Fontanetti Christofoletti / Banca: Maria do Carmo Pimentel Batitucci / Mestre