Própolis: caracterização físico-química, atividade antimicrobiana e antioxidante
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2009-09-21
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Universidade Federal de Viçosa
Resumo
A própolis é um produto natural, produzido pelas abelhas que coletam resina de brotos, folhas e exsudatos de plantas e é modificado pela mistura de secreções salivares, pólen e cera. Destaca-se por suas inúmeras propriedades, dentre as quais estão: antimicrobiana, anti-inflamatória, cicatrizante, antioxidante, antitumoral, anestésica e anticariogênica. Inúmeros fatores contribuem para a diversidade de características e qualidade da própolis, tornando-a um material de composição complexa. Diversos autores conferem ao extrato aquoso as mesmas propriedades do extrato etanólico. Devido à discrepância com relação às informações sobre as propriedades do extrato aquoso, faz-se necessário o desenvolvimento de estudos comparativos visando esclarecer tais aspectos. Assim, o presente trabalho teve por objetivo determinar alguns indicadores de qualidade de extratos de própolis realizando quatro experimentos. O primeiro experimento teve como objetivo avaliar a atividade antifúngica de extratos aquoso e etanólico de própolis provenientes de duas amostras distintas (própolis C e V) e obtidos a partir de três temperaturas de extração (50, 70 e 95ºC) e com duas concentrações finais (2 e 4%) sobre alguns fungos (Alternaria brassicae, Botrytis cinerea, Colletotrichum gloeosporioides, Colletotrichum musae, Lasiodiplodia theobromae, Mucor sp., Phythophtora capsici, Rhizoctonia solani, Rhizopus stolonifer e Sclerotium rolfsii) causadores de doenças pós-colheita. Os extratos etanólicos apresentaram maior capacidade de inibição fúngica, enquanto os extratos aquosos foram pouco eficazes. Dentre os efeitos principais testados, o solvente mostrou grande importância na extração dos compostos antifúngicos, onde os extratos etanólicos de própolis foram os mais efetivos. A concentração de 4% foi a que apresentou melhor desempenho no controle do crescimento micelial. De modo geral, o comportamento foi variável entre as diferentes espécies de fungos. Não foi constatada associação ao grupo filogenético dos fungos e a sua sensibilidade a própolis. O fungo mais sensível foi Mucor sp., com 100% de inibição do crescimento micelial pelos extratos etanólicos. Lasiodiplodia theobromae foi o fungo mais tolerante aos extratos de própolis. O segundo experimento teve como objetivo avaliar a atividade antibacteriana de diferentes extratos etanólico e aquoso de própolis, oriundos de duas origens distintas (própolis C e V) e obtidos a partir de duas temperaturas (50 e 70ºC) de extração sobre três isolados bacterianos (Staphylococcus aureus, Escherichia coli e Salmonella choleraesius). Verificou-se que os dez extratos de própolis estudados tiveram pouco efeito sobre a inibição do crescimento de E. coli (halos de 0 a 1,2 mm). Para S. aureus os extratos etanólicos preparados a 70ºC, de ambas as origens apresentaram um halo de inibição considerável (2,3 mm). Em relação à S. choleraesius os extratos etanólico a 70ºC (própolis C), aquoso a 50ºC (própolis V) e etanólico a 70ºC (própolis V) apresentaram uma boa atividade antibacteriana com halos que variaram de 2,0 a 3,1 mm. O terceiro experimento teve como objetivo realizar a caracterização físico-química (umidade, cinzas, fenóis totais, flavonóides totais, compostos voláteis, espectro de absorção), análise microbiológica (contagem de fungos filamentosos e leveduras), caracterização sensorial e atividade antioxidante da própolis bruta e dos extratos aquosos e etanólicos de própolis. As duas própolis brutas (C e V) estudadas estão de acordo com o exigido pela legislação quanto ao percentual de cinzas, com valores de 2,6 e 3,4%. Quanto à umidade a própolis V encontra-se dentro do padrão exigido 7,6% (máximo de 8%), enquanto a própolis C apresentou um valor acima do permitido, 8,6%. O percentual de compostos voláteis foi de 0,63% para a própolis C e de 1,0% para a amostra de própolis V. No espectro de absorção, os extratos apresentaram uma variação de 290,6 a 299,2 nm, sendo que os etanólicos foram os que apresentaram os maiores comprimentos de onda. A própolis C e extratos etanólico e aquoso apresentaram contagem baixa para fungos filamentosos e levedura (< 10 UFC/g), e a própols V uma contagem de 4,9 x103 UFC/g. Quanto a caracterização sensorial, as amostras de própolis brutas apresentaram conformidade com a legislação brasileira vigente. Em relação ao teor de fenóis, todos os extratos avaliados encontram-se de acordo com a legislação brasileira. Os extratos aquosos apresentaram teores de flavonóides que variaram de 0,21 a 0,30% e os extratos etanólicos valores de2,66 a 3,60%. Em relação a atividade antioxidante observou-se que todos os extratos apresentaram uma baixa atividade, sendo que os extratos etanólicos com maior atividade antioxidante em relação aos extratos aquosos. No quarto ensaio realizaram-se dois experimentos com o objetivo de avaliar o efeito inibitório de diferentes extratos comerciais de própolis (aquoso, etanólico e propilenoglicol) nas concentrações 0,1; 0,5; 1,0 e 1,5% sobre a germinação de conídios dos fungos Aspergillus flavus, Alternaria brassicae, Bipolaris sorokiniana e Colletotrichum musae, e o crescimento micelial dos fungos Botrytis cinerea, Colletotrichum gloeosporioides, Lasiodiplodia theobromae, Phythophthora capsici e Rhizopus stolonifer, em que utilizaram-se preparações de extratos de própolis (aquoso e alcoólico) obtidas no laboratório, nas concentrações de 2 e 3%. Todos os fungos estudados são considerados importantes fitopatógenos em pré e pós-colheita. Os extratos aquosos não apresentaram efeito inibitório da germinação e do crescimento micelial dos fungos avaliados. Os extratos alcoólicos e de propilenoglicol inibiram em mais de 80% a germinação de conídios de A. brassicae e C. musae nas concentrações de 0,5 e 1,0%, respectivamente. Embora a germinação dos conídios de A. flavus e B. sorokiniana não tenha sido inibida, observou-se a redução do tubo germinativo em relação à testemunha. Maiores índices de inibição do crescimento micelial foram alcançados nas preparações com álcool a 3% para todos os fungos testados. Lasiodiplodia theobromae foi o fungo menos suscetível, enquanto P. capsici o mais suscetível. Os demais fungos apresentaram um comportamento variável.
Propolis is a natural product, made by bees, collected from buds, leaves and plant exudates and modified mixing salivary secretions, pollen and waxes. Reaches a high position, due to its several properties: antimicrobial, antinflamathory, healing, antioxidant, antitumoral, anesthetic and anticariogenic. Several factors contribute to the diversity of characteristics and quality of the propolis, turning it a material of complex composition. Due to divergences in relation of the information about propriety of the aqueous extract, is necessary the development of the comparative studies aiming to explain this aspects. Thus, the present study aimed to evaluate some quality indicators of aqueous and ethanolic extract realizing four experimental assays. The first assay had as objective evaluates the antifungal activity of aqueous and ethanolic extracts obtained from three extraction temperatures (50, 70 and 95 oC) and two final concentrations of the extracts added to the growth medium (2 and 4%) against some fungi (Alternaria brassicae, Botrytis cinerea, Colletotrichum gloeosporioides, Colletotrichum musae, Lasiodiplodia theobromae, Mucor sp., Phythophtora capsici, Rhizoctonia solani, Rhizopus stolonifer e Sclerotium rolfsii ) causers of the post-harvest diseases in fruits and vegetables. Ethanolic extracts presented higher capacity of fungal inhibition, while aqueous extracts were less effective. Among the main effects tested the solvent showed great importance in the extraction of the antifungal compounds, showing that propolis ethanolic extracts were more effective, while aqueous extract were almost ineffective despite of the fungal species tested. The 4% concentration presented better performance in the control of mycelial growth of all tested fungi. The propolis extracts showed a variable effect among the different fungi species. There was no association between the phylogenetic fungi group and its sensibility to propolis. The most susceptible fungus was Mucor sp., with 100% of inhibition of the growth mycelial for the ethanolic extracts. Lasiodiplodia theobromae was the most tolerant fungus. It can be concluded that ethanolic extract of propolis possess a great potential as alternative control to fungi that causes of post-harvest disease, while aqueous extracts still need to be better evaluated. The second assay aimed to evaluate the antibacterial activity of ethanolic and aqueous propolis extracts obtained from two distinct origins (namely propolis V and C) with three temperatures of extraction (50, 70 and 95 oC, except for alcohol extracts that were obtained only at 50 and 70 oC) against three bacterial strains, one of each of the following species: Staphylococcus aureus, Escherichia coli and Salmonella choleraesius. It was verified that the ten extracts obtained in this study had little effect on the growth inhibition of E. coli (halos of 0 to 1.2 mm). The ethanolic extracts prepared at 70ºC from both origins presented a considerable inhibition (2,3 mm) against S. aureus. For S. choleraesius the ethanolic extracts at 70 ºC (propolis C), aqueous at 50 ºC (propolis V), ethanolic at 70 ºC (propolis V) presented a good antibacterial activity with halos that varied of 2.0 to 3.1 mm. The third assay aimed at study the physiochemical characterization (ashes; lost weight on drying (humidity); total phenols and total flavonoids of the crude samples; volatile compounds contents; spectrum of absorption); microbiological analysis (counting of molds and yeasts); sensory analysis and antioxidant activity from two distinct propolis, marketable in Minas Gerais. Both samples, V and C of crude propolis are in agreement with Brazilian regulation as for the percentile of ashes, (2,6 and 3,4%). Moisture of the sample V was 7.6% (maximum of 8%), however the sample C presented value of 8.6%, which is above of maximum allowed. Volatile compounds content was 0.63% on propolis C and 1.0% for propolis V. As for of the absorption spectrum, the extracts ranged from 290.6 to 299.2 nm, and the ethanolic extracts presented the highest wavelengths. Propolis C and ethanolic and aqueous extracts presented low counting for molds and yeast (<10 UFC.g-1), while crude sample V presented a counting of 4.9 x103 UFC.g-1. Regarding the sensory characterization, both samples of crude propolis were in conformity with the limits established by the Brazilian regulation. All extracts were low in antioxidant activity, though, ethanolic extracts presented higher activity than the aqueous ones. In the fourth assay, two experiments were carried out to determine inhibitory effects of propolis extract against some fungi. The first one evaluated the effect of the different commercial extracts of propolis (aqueous, ethanolic and propylene glycol) in the concentrations 0.1; 0.5; 1.0 and 1.5% against conidia germination of Aspergillus flavus, Alternaria brassicae, Bipolaris sorokiniana and Colletotrichum musae. The second one evaluated the mycelial growth of Botrytis cinerea, Colletotrichum gloeosporioides, Lasiodiplodia theobromae, Phythophthora capsici e Rhizopus stolonifer, with laboratory propolis preparations (aqueous and ethanolic) in the final concentrations of 2 and 3% on the growth medium. The nine fungi studied are considered important pre and post-harvest pathogens. In the first assay, the aqueous extracts had no inhibitory effect on conidia germination and mycelial growth of tested fungi. Commercial ethanolic and propylene glycol extracts showed inhibitory effect over 80% on the conidia germination of Alternaria brassicae and Colletotrichum musae, even when used the lowest concentration. Although conidia germination of Aspergillus flavus and Bipolaris sorokiniana was not inhibited, its germ tubes were clearly shorter than the controls (no propolis addition to growth medium). Regarding the mycelial growth assay, the higher inhibitions were achieved with ethanol preparations added to the medium at 3% to all test fungi. Lasiodiplodia theobromae was the less susceptible fungus, while Phythophtora capsici was the most susceptible. Botrytis cinerea, Colletotrichum gloeosporioides and Rhizopus stolonifer showed a variable behavior.
Propolis is a natural product, made by bees, collected from buds, leaves and plant exudates and modified mixing salivary secretions, pollen and waxes. Reaches a high position, due to its several properties: antimicrobial, antinflamathory, healing, antioxidant, antitumoral, anesthetic and anticariogenic. Several factors contribute to the diversity of characteristics and quality of the propolis, turning it a material of complex composition. Due to divergences in relation of the information about propriety of the aqueous extract, is necessary the development of the comparative studies aiming to explain this aspects. Thus, the present study aimed to evaluate some quality indicators of aqueous and ethanolic extract realizing four experimental assays. The first assay had as objective evaluates the antifungal activity of aqueous and ethanolic extracts obtained from three extraction temperatures (50, 70 and 95 oC) and two final concentrations of the extracts added to the growth medium (2 and 4%) against some fungi (Alternaria brassicae, Botrytis cinerea, Colletotrichum gloeosporioides, Colletotrichum musae, Lasiodiplodia theobromae, Mucor sp., Phythophtora capsici, Rhizoctonia solani, Rhizopus stolonifer e Sclerotium rolfsii ) causers of the post-harvest diseases in fruits and vegetables. Ethanolic extracts presented higher capacity of fungal inhibition, while aqueous extracts were less effective. Among the main effects tested the solvent showed great importance in the extraction of the antifungal compounds, showing that propolis ethanolic extracts were more effective, while aqueous extract were almost ineffective despite of the fungal species tested. The 4% concentration presented better performance in the control of mycelial growth of all tested fungi. The propolis extracts showed a variable effect among the different fungi species. There was no association between the phylogenetic fungi group and its sensibility to propolis. The most susceptible fungus was Mucor sp., with 100% of inhibition of the growth mycelial for the ethanolic extracts. Lasiodiplodia theobromae was the most tolerant fungus. It can be concluded that ethanolic extract of propolis possess a great potential as alternative control to fungi that causes of post-harvest disease, while aqueous extracts still need to be better evaluated. The second assay aimed to evaluate the antibacterial activity of ethanolic and aqueous propolis extracts obtained from two distinct origins (namely propolis V and C) with three temperatures of extraction (50, 70 and 95 oC, except for alcohol extracts that were obtained only at 50 and 70 oC) against three bacterial strains, one of each of the following species: Staphylococcus aureus, Escherichia coli and Salmonella choleraesius. It was verified that the ten extracts obtained in this study had little effect on the growth inhibition of E. coli (halos of 0 to 1.2 mm). The ethanolic extracts prepared at 70ºC from both origins presented a considerable inhibition (2,3 mm) against S. aureus. For S. choleraesius the ethanolic extracts at 70 ºC (propolis C), aqueous at 50 ºC (propolis V), ethanolic at 70 ºC (propolis V) presented a good antibacterial activity with halos that varied of 2.0 to 3.1 mm. The third assay aimed at study the physiochemical characterization (ashes; lost weight on drying (humidity); total phenols and total flavonoids of the crude samples; volatile compounds contents; spectrum of absorption); microbiological analysis (counting of molds and yeasts); sensory analysis and antioxidant activity from two distinct propolis, marketable in Minas Gerais. Both samples, V and C of crude propolis are in agreement with Brazilian regulation as for the percentile of ashes, (2,6 and 3,4%). Moisture of the sample V was 7.6% (maximum of 8%), however the sample C presented value of 8.6%, which is above of maximum allowed. Volatile compounds content was 0.63% on propolis C and 1.0% for propolis V. As for of the absorption spectrum, the extracts ranged from 290.6 to 299.2 nm, and the ethanolic extracts presented the highest wavelengths. Propolis C and ethanolic and aqueous extracts presented low counting for molds and yeast (<10 UFC.g-1), while crude sample V presented a counting of 4.9 x103 UFC.g-1. Regarding the sensory characterization, both samples of crude propolis were in conformity with the limits established by the Brazilian regulation. All extracts were low in antioxidant activity, though, ethanolic extracts presented higher activity than the aqueous ones. In the fourth assay, two experiments were carried out to determine inhibitory effects of propolis extract against some fungi. The first one evaluated the effect of the different commercial extracts of propolis (aqueous, ethanolic and propylene glycol) in the concentrations 0.1; 0.5; 1.0 and 1.5% against conidia germination of Aspergillus flavus, Alternaria brassicae, Bipolaris sorokiniana and Colletotrichum musae. The second one evaluated the mycelial growth of Botrytis cinerea, Colletotrichum gloeosporioides, Lasiodiplodia theobromae, Phythophthora capsici e Rhizopus stolonifer, with laboratory propolis preparations (aqueous and ethanolic) in the final concentrations of 2 and 3% on the growth medium. The nine fungi studied are considered important pre and post-harvest pathogens. In the first assay, the aqueous extracts had no inhibitory effect on conidia germination and mycelial growth of tested fungi. Commercial ethanolic and propylene glycol extracts showed inhibitory effect over 80% on the conidia germination of Alternaria brassicae and Colletotrichum musae, even when used the lowest concentration. Although conidia germination of Aspergillus flavus and Bipolaris sorokiniana was not inhibited, its germ tubes were clearly shorter than the controls (no propolis addition to growth medium). Regarding the mycelial growth assay, the higher inhibitions were achieved with ethanol preparations added to the medium at 3% to all test fungi. Lasiodiplodia theobromae was the less susceptible fungus, while Phythophtora capsici was the most susceptible. Botrytis cinerea, Colletotrichum gloeosporioides and Rhizopus stolonifer showed a variable behavior.
Descrição
Palavras-chave
Própolis, Qualidade, Propriedades, Propolis, Quality, Properties
Citação
SILVA, Aline Fonseca da. Propolis: physiochemical characterization, antifungal activity and antioxidant. 2009. 145 f. Tese (Doutorado em Ciência de Alimentos; Tecnologia de Alimentos; Engenharia de Alimentos) - Universidade Federal de Viçosa, Viçosa, 2009.