ABSTRACT
Aims: Bark, leaves and gum of cashew (Anacardium occidentale L.) have been reported to be effective in curtailing the growing problems of resistance of bacterial pathogens. The in vitro activity of aqueous extracts of cashew apple peels was determined in this study against two clinically important pathogens. Place and Duration of Study: The work was conducted at the Department of Microbiology, Ekiti State University, Ado-Ekiti, Nigeria and processed immediately. This study was carried out between June, 2009 and January, 2010. Methodology: Bioautographic method was used to test the antibacterial activity of aqueous (cold and hot water) extracts of cashew apple peels on Escherichia coli O157:H7 and methicillin-resistant Staphylococcus aureus (MRSA). The zones of inhibition of the extracts were compared. Results: The activity of the fifth hour hot water extract was highest with zones of inhibition of 415.48 and 346.30 sq. mm against E. coli O157:H7 and MRSA respectively. E. coli O157:H7 was more susceptible to the extract with the zone of inhibition ranging between 176.79 and 283.53 sq. mm while that of MRSA was153.94 - 346.30 sq. mm. The 5 h extract of cold water was more potent on the test organisms with 615.75 and 490.87 sq. mm diameters of inhibition on E. coli O157:H7 and MRSA respectively. Cold water extracts produced more active compounds (13 biologically active spots) that inhibited the growth of the test organisms than the hot water extracts, with six spots. Conclusion: The extracts of the peels of the cashew apple against the test organisms are promising. However, the nature and mechanisms of action of the biologically active compounds in the extracts are still open to investigation.
ABSTRACT
Leaf oil prepared by hydrodistillation of Alpinia zerumbet Pers.) B.L. Burtt & R.M. Sm. was analyzed by GC/FID and GC/MS to determine the major compounds and it was also evaluated for antimicrobial activity. The oil presented a high content of oxygenated monoterpenes (52.5 percent), terpinen-4-ol, 1,8 cineole and γ-terpinene as the major constituents. The antimicrobial activity of leaf oil was tested by drop diffusion and bioauthography methods. Through drop test, all bacteria and fungi tested were inhibited by leaf oil. Preparative TLC (thin-layer chromatography) plates were developed using the mobile phase hexane: ethyl acetate (70:20, v/v) and fractions of the leaf oil were separated in three zones, scraped, extracted from silica and identified by GC/MS. The bioauthography method permitted to verify pronounced inhibition of Cryptococcus neoformans by fractions F2 and F3, both rich in oxygenated monoterpenes. The fraction F2 comprised 1,8 cineole (9.6 percent), linalool (3.7 percent) and caryophyllene oxide (5.4 percent), while fraction F3 showed mainly terpinen-4-ol (43.6 percent) and an amount of 32.7 percent oxygenated sesquiterpenes. The obtained data may be used to suggest the constituents of A. zerumbet leaf oil involved in antimicrobial activity.
O óleo essencial de folhas de Alpinia zerumbet (Pers.) B.L. Burtt & R.M. Sm., extraído por hidrodestilação, foi analisado por CG/DIC e CG/EM a fim de se determinar sua composição e atividade antimicrobiana. O óleo apresentou um elevado teor de monoterpenos oxigenados (52,5 por cento): terpinen-4-ol, 1,8-cineol e γ-terpineno como principais constituintes. A atividade antimicrobiana do óleo foi analisada pelos testes de difusão em agar e ensaios de bioautografia. Através do teste de difusão em agar, todas as bactérias e fungos testados foram inibidos pelo óleo essencial. Placas preparativas de CCD (cromatografia em camada fina) foram eluídas em hexano: acetato de etila (70:20, v/v), e o óleo foi separado em três frações que foram raspadas, extraídas da sílica e identificadas por CG/EM. A análise bioautográfica permitiu detectar pronunciada inibição de Cryptococcus neoformans pelas frações F2 e F3, ambas ricas em monoterpenos oxigenados. A fração F2 apresentou 1,8 cineol (9,6 por cento), linalol (3,7 por cento) e óxido de cariofileno (5,4 por cento), enquanto a fração F3 mostrou principalmente terpinen-4-ol (43,6 por cento) e cerca de 32,7 por cento de sesquiterpenos oxigenados. Os dados obtidos indicam os componentes do óleo de A. zerumbet envolvidos na atividade antimicrobiana.
ABSTRACT
Brazilian propolis from São Paulo state was submitted to extraction using several solvents, resulting in extracts with different composition. These extracts were submitted to Thin Layer Chromatography (TLC). Bioauthographic analysis of the TLC plates identified fractions with inhibitory activity, which were then analysed by High Performance Liquid Chromatography (HPLC). In vitro assays, commonly used to evaluate the activity of propolis against Gram-positive bacteria, were compared to determine which rendered the most consistent results. The bactericidal activity of these extracts were analysed by Serial Dilution in Tubes and Agar Plate Diffusion. Serial Dilution in Tubes obtained the most consistent results, with the Minimal Bactericidal Concentration of the extracts ranging between 2.5 and 20.0 mg/mL, for the species of Gram-positive bacteria tested. The results of the Agar Plate Diffusion were directly proportional to the hydro-solubility of the extracts and did not evaluate their bactericidal activity correctly. The bactericidal activity of this sample of propolis was due to the combined effect of several components that were identified by HPLC and were best extracted using 50% ethanol as a solvent.
Própolis brasileira, proveniente do estado de São Paulo, foi submetida à extração usando vários solventes, resultando em extratos com diferentes composições. Estes extratos foram submetidos à Cromatografia em Camada Delgada (CCD). Análise bioautográfica das placas de CCD permitiu identificar as frações com atividade antimicrobiana, que foram então analisadas por Cromatografia Líquida de Alta Eficiência (CLAE). Ensaios in vitro freqüentemente utilizados para avaliar a atividade de própolis frente a bactérias Gram-positivas foram comparados para determinar qual renderia os resultados mais consistentes. A atividade bactericida destes extratos foi analisada por Diluição Seriada em Tubos e por testes de Difusão em Agar. O método de Diluição em Tubos permitiu obter os resultados mais consistentes e a Concentração Bactericida Mínima dos extratos variou entre 2,5 e 20,0 mg/mL, para as espécies de bactérias Gram-positivas testadas. Os resultados do método de Difusão em Agar foram diretamente proporcionais à hidrossolubilidade dos extratos, e não avaliaram a atividade bactericida corretamente. A atividade bactericida desta amostra resultou da combinação de vários componentes, identificados por CLAE, que foram extraídos preferencialmente usando etanol 50% como solvente.