ABSTRACT
There is a growing need to discover and develop alternative therapies for the treatment of mastitis caused by Staphylococcus spp. and multidrug-resistant bacterial infections. This study examined the chemical composition and antimicrobial potential of two propolis extracts (EPA and EPB) against seventy-seven isolates of Staphylococcus spp. obtained from subclinical bovine mastitis; three clinical strains of MRSA and two from clinical strains of S. aureus ATCC, identified as S. aureus ATCC 6538 and S. aureus ATCC 25923. The total phenolic content was determined by the Folin-Ciocalteau method, the total flavonoid content by the Dowd method and the phenolic profile was quantified by HPLC-DAD. The MBC values of the extracts were evaluated by broth microdilution method. The amount of total phenolic and flavonoid compounds was higher in EPA than EPB. Both extracts revealed the presence of caffeic, coumaric, cinnamic, ferulic and 3,4-dihydroxybenzoic acids, with higher concentrations of coumaric and cinnamic acids. Staphylococcus spp. isolates were susceptible to EPA (90.9%), EPB (83.1%) and oxacillin (80.5%). The oxacillin susceptible isolates were also susceptible to EPA (70.1%) and EPB (80.6%), whereas those oxacillin-resistant strains were also susceptible to EPA (40.0%) and to EPB (26.7%). MBC ranged from 34.3 to 68.7µm/mL for EPA and from 68.7 to 137.5µg/mL for EPB. Both extracts inhibited significantly (100%) the clinical strains of MRSA, S. aureus ATCC 6538 and S. aureus ATCC 25923 at the concentration of 68.7µg/mL. It is concluded that both extracts of propolis, whose main constituents are coumaric and cinnamic acids, have high antimicrobial activity against the microorganisms studied, and EPA also against oxacillin-resistant strains. These findings reinforce its potential use for the treatment of bovine mastitis.(AU)
É cada vez mais oportuna a necessidade de descobrir e desenvolver terapias alternativas para tratamento da mastite causada por Staphylococcus spp. e de infecções bacterianas multirresistentes. Este estudo examinou a composição química e o potencial antimicrobiano de dois extratos etanólicos de própolis (EPA e EPB) contra setenta e sete isolados de Staphylococcus spp. obtidos a partir de mastite bovina subclínica; três estirpes clínicas de MRSA e duas de linhagens clínicas de S. aureus ATCC, identificadas como, S. aureus ATCC 6538 e S. aureus ATCC 25923, ambas metacilina resistentes. O teor total de fenólicos foi determinado pelo método de Folin-Ciocalteau, o teor de flavonoides totais pelo método Dowd e o perfil fenólico foi quantificado por HPLC-DAD. CBM dos extratos foi avaliada pelo método de microdiluição em caldo. A quantidade total de compostos fenólicos e flavonoides foi maior no EPA do que no EPB. Ambos os extratos revelaram a presença dos ácidos cafeico, cumárico, cinâmico, ferúlico e 3,4-di-hidroxibenzóico, com maiores concentrações de ácidos cumárico e cinâmico. Os isolados de Staphylococcus spp. foram sensíveis a EPA (90,9%), EPB (83,1%) e oxacilina (80,5%). Os isolados suscetíveis à oxacilina também foram suscetíveis ao EPA (70,1%) e ao EPB (80,6%), enquanto os do resistente à oxacilina foram suscetíveis ao EPA (40,0%) e ao EPB (26,7%). MBC variou de 34,3 a 68,7µm/mL para EPA e de 68,7 a 137,5µg/mL para EPB. Ambos os extratos inibiram significativamente (100%) as linhagens clínicas de MRSA, S. aureus ATCC 6538 e S. aureus ATCC 25923 na concentração de 68,7µg/mL. Conclui-se que os extratos etanólicos da própolis, cujos principais constituintes são os ácidos cumário e cinâmico, possuem atividade antimicrobiana contra os micro-organismos estudados, e o EPA também contra as cepas resistentes à oxacilina. Estes achados reforçam seu potencial uso para o tratamento da mastite bovina.(AU)
Subject(s)
Oxacillin , Propolis/immunology , Staphylococcus , Drug Resistance, Multiple, Bacterial , Phenolic Compounds/analysisABSTRACT
Própolis é composta por substâncias resinosas, coletadas de várias plantas por abelhas africanizadas, adicionando pólen, cera e secreções glandulares, enquanto para a produção de geoprópolis (Geo) as abelhas sem ferrão utilizam materiais resinosos de plantas, cera, e ainda acrescentam terra ou barro. As atividades biológicas da própolis produzida por Apis mellifera têm sido extensivamente estudadas, mas ainda são escassos os trabalhos sobre as propriedades da geoprópolis. Assim, o presente trabalho se propôs a investigar as atividades citotóxica, imunomoduladora, antibacteriana e antifúngica do extrato hidroalcoólico de geoprópolis de Melipona fasciculata Smith. Foi realizada a caracterização química da Geo, verificando que o principal grupo de compostos encontrados foram triterpenos. No tocante às células do carcinoma da laringe humana (HEp-2), Geo apresentou ação inibitória em concentrações mais elevadas; entretanto, esta ação foi somente citostática. Geo apresentou ação imunomoduladora sobre monócitos humanos, estimulando a produção das citocinas estudadas (TNF-α e IL-10). Não foi observada ação antibacteriana contra S. aureus e E. coli, havendo inibição destas linhagens somente com altas concentrações do extrato. Contudo, a associação de Geo com antibióticos favoreceu a ação do cloranfenicol sobre S. aureus. A Geo exerceu ação inibitória sobre Pythium insidiosum, tanto para o isolado obtido de caso humano como de eqüinos. Os resultados sugerem que a Geo apresentou inúmeras propriedades biológicas; contudo, sua ação é menor em comparação à própolis produzida por abelhas africanizadas, talvez devido ao baixo rendimento do extrato e sua difícil solubilidade...
Propolis is composed of resinous substances collected from various plants by Africanized honeybees, adding polen, wax and bee secretions, while for geopropolis (Geo) production stingless bees use resinous materials from plants, wax, but also adding mud or clay. The biological activities of propolis produced by Apis mellifera have been extensively studied, but there are few studies on Geo properties. Thus, the present study investigated the cytotoxic, immunomodulatory, antibacterial and antifungal activities of hydroalcoholic extract of Geo produced by Melipona fasciculata Smith. The chemical characterization of Geo was performed, revealing that the main group of compounds was triterpenes. Geo exerted an inhibitory action on human larynx carcinoma cells (HEp-2) using high concentrations, however, it was only a cytostatic action. Geo exhibited immunomodulatory effects on human monocytes, stimulating cytokines production (TNF-α and IL-10). No antibacterial activity was seen against S. aureus and E. coli. However, the combination of Geo with chloramphenicol favored its effect against S. aureus. Geo exerted an inhibitory action against Pythium insidiosum, both for human or equine isolates. Data suggested that Geo presented several properties; however, its activity is lower compared to propolis produced by honeybees, perhaps due to the low yield of the extract and its difficult solubility...
Subject(s)
Anti-Infective Agents , Bees , Propolis/immunologyABSTRACT
The activity of 23 samples of ethanolic brown propolis, from the State of Mato Grosso, was investigated against Escherichia coli ATCC 25922. The values of physical and chemical parameters showed significant variation among samples. The percentage of dry extract ranged from 2.6 to 27.6 percent. The index of oxidation varied from 3 to 519 seconds. All samples showed the percentage of wax higher than the limit preconized by the legislation, with values varying from 3.4 to 74.6 percent. The quantification of phenolic and flavonoid compounds, responsible for antimicrobial activity, ranged from 0.1 to 5.0 (w/w) and 0.02 to 0.66 (w/w), respectively, being that the higher the index of phenolic compounds the larger the zones of inhibition. Antibacterial activity was observed in seven out of the 23 samples, demonstrating zones of inhibition ranging from 10 to 11.3mm. For these active samples, the minimum inhibitory concentration was determined, ranging from 125 to 1000mg/mL. The value of MIC in 42.9 percent of these samples was 250mg/mL. These results contribute to the establishment of physical and chemical parameters for the regulation of brown propolis and indicate possible therapeutic applicability in the development of formulations for the treatment of infections caused by E. coli.
Subject(s)
Animals , Anti-Bacterial Agents , Escherichia coli , Propolis/pharmacology , Propolis/immunology , Propolis/therapeutic use , Anti-Infective Agents , Biological Assay/methodsABSTRACT
Propolis is a resinous bee product with an immuno-regulatory effect. It is superior to many other natural agents as its pharmacological and biological activity is known, especially after fractionation and characterization of its individual components. The ability of propolis to enhance cytokine response in mice infected with Trichinella spiralis larvae assessed. An enzyme linked immunosorbant assay [ELISA] was performed on cell culture supernatants of mesenteric lymph nodes for in vitro analysis of interleukin [IL-4] and interferon gamma [IFN-delta] on day 7 and 14 post infection. Also the serum level of NO [nitric oxide] was investigated on the same days. Supernatants and sera were harvested from separate groups of infected mice receiving propolis orally either pre-infection or on the same day of infection in low and high doses; 100 or 200 mg/kg for five consecutive days. Propolis treated mice showed aTh1/Th-2 mixed cytokine pattern yet polarized to Th-2 response with high IL-4 secretion on days [7,14]. The nitric oxide level showed non significant increase in treated mice versus control infected mice. Propolis was found to be effective in accelerating the worm expulsion with a percent reduction of adult worms in the intestine varied between 68.8% to 73.9% when administered before infection.And 72% to 74.4% when administered post infection in 100 mg/kg or 200 mg/kg respectively on day 7 compared to the control group. However the maximum reduction occurred on day 14 as it reaches up to 95.5% in mice treated with 200 mg/kg post infection. This effect was not time or dose dependant.Concerning the histopathological examination, propolis treated mice showed increase in intestinal villus height with marked improvement of the villus architecture and marked infiltration of mononuclear cells/ crypt unit compared to control group. The obtained results indicated that propolis causes augmented immune responses shifted to T-helper type 2 cytokine response that showed a protective effect against local intestinal T. spiralis infection in mice