ABSTRACT
A total of 22 endophytic fungi isolated from coffee (Coffea arabica L.) were cultivated in vitro and their crude extracts tested. The screening was carried out using the agar diffusion method against Staphylococcus aureus, Escherichia coli and Candida albicans. The most effective isolate was Alternaria alternata, and subsequently, its extract was assayed. The total phenolic content was 3.44 μg GAE/mg of the crude extract. For the antibacterial and antifungal activity assays, minimum inhibitory concentrations (MIC) and minimum bactericidal and fungicidal concentrations (MBC and MFC) were determined. The ranges of MIC values were 50-100 μg/mL for S. aureus and 400-800 μg/mL for E. coli. The extract did not show activity in the tested concentrations for C. albicans. The fungal crude extract was assayed for antioxidant activities. Its ability to scavenge DPPH radicals and antioxidant activity by β-carotene/linoleic acid system oxidation was not significant. In addition, antitumor activity was studied using the MTT assay. At a dilution of 400 μg/mL, the extract displayed a cytotoxic activity of approximately 50 percent towards HeLa cells in vitro. The results indicate that endophytic fungi could be a promising source of bioactive compounds and warrant further study.
Total de 22 fungos endofíticos isolados de café (Coffea arabica L.) foi cultivado in vitro e seus extratos testados. A triagem foi conduzida pelo método de difusão em agar contra bactérias Gram-positiva, Gram-negativa e uma levedura. O isolado mais efetivo foi Alternaria alternata e, subsequentemente, seu extrato foi analisado. O conteúdo de fenólicos totais do extrato bruto foi de 3,44 μg EAG/mg de extrato. Para os testes de atividade antimicrobiana, a concentração inibitória mínima (CIM) e concentração bactericida e fungicida mínima (CBM e CFM) contra Staphylococcus aureus, Escherichia coli e Candida albicans foram determinadas. Resultados da CIM variaram entre 50-100 μg/mL para S. aureus e 400-800 μg/mL para E. coli. O extrato bruto não apresentou atividade nas concentrações testadas para C. albicans. Foram analisadas as atividades antioxidantes do extrato bruto. Sua habilidade para seqüestrar radicais DPPH e a atividade antioxidante pela oxidação do sistema β-caroteno/ácido linoléico não foram significativas. Além disso, a atividade antitumoral foi estudada pelo teste do MTT. À diluição de 400 μg/mL, o extrato apresentou atividade de aproximadamente 50 por cento sobre as células HeLa in vitro. Os resultados indicam que fungos endófitos poderiam ser uma fonte promissora de compostos bioativos necessitando de estudos futuros.
Subject(s)
Alternaria/growth & development , Alternaria/chemistry , Coffea Cruda/analysis , Fermentation , Antioxidants/metabolism , Chemical Compounds/analysis , Drug SynergismABSTRACT
The morphological features of a Penicillium, isolated from Brazilian cerrado soil, were characterized and showed to be distinctly different from all well-defined Penicillium species. Chemical and biological investigation on the ethyl acetate extract of this Penicillium isolate resulted in the isolation of three new naphthalenoids: a major metabolite, methyl 6-acetyl-4-methoxy-5,7,8-trihydroxynaphthalene-2-carboxylate and two minor ones, methyl 6-acetyl-4-methoxy-7,8-dihydroxynaphthalene-2-carboxylate and methyl 6-acetyl-4-methoxy-5,8-dihydroxynaphthalene-2-carboxylate. Their structures were determined based on their mono and bidimensional nuclear magnetic resonance data. Acetyl, allyl and methoxyl derivatives of the major metabolite were prepared in order to establish structure-activity relation. Antimicrobial activity of the major natural product and its semi-synthetic derivatives was screened by macro dilution methodology and the corresponding minimum inhibitory concentrations were determined. Natural secondary metabolite methyl 6-acetyl-4-methoxy-5,7,8-trihydroxynaphthalene-2-carboxylate, isolated in a very high yield (0.3175 mg.L-1) showed to be the most active compound, possessing expressive activity against Candida albicans (minimum inhibitory concentration (MIC) 32 ug/mL), Listeria monocitogenes and Bacillus cereus (MIC 64 µg/mL for both).
Subject(s)
Animals , Fungi/isolation & purification , Penicillium/isolation & purification , Penicillium/classification , Penicillium/metabolism , Brazil , Magnetic Resonance Spectroscopy/methods , Methylation , Disk Diffusion Antimicrobial Tests/methodsABSTRACT
A obtenção de protoplastos é uma ferramenta importante para a transformação genética de fungos. Neste trabalho foi estudada a influência de fatores como idade micelial, tipo e concentração de enzimas e estabilizadores osmóticos na produção de protoplastos de Aspergillus ochraceus. Os melhores resultados de produção de protoplastos foram obtidos utilizando-se NH4Cl 0,8mol L-1 como estabilizador osmótico, micélio com 24h de crescimento e a combinação de Lysing Enzymes e Meicelase ambas, a 20mg mL-1. Entretanto, bons resultados foram também obtidos com a utilização apenas de Lysing Enzymes.
Production of protoplasts is an important tool for genetic transformation of fungi. A protocol for protoplasts production in Aspergillus ochraceus was developed, evaluating culture aging of mycelium, different commercial enzymes and osmotic stabilizers. The best results were obtained with NH4Cl 0.8mol L-1 as osmotic stabilizer, mycelial age of 24 hours and Lysing Enzymes (20mg mL-1) plus Meicelase (20mg mL-1) as lytic enzymes. Good results were also obtained with Lysing Enzymes alone.
ABSTRACT
Visando conhecer a diversidade e a incidência de fungos emboloradores e manchadores de madeira da região Amazônica, foi realizado um levantamento em 12 espécies florestais, estocadas em quatro indústrias madeireiras de Manaus. De cada espécie florestal selecionaram-se cinco toras, das quais retiraram-se amostras de onde procedeu-se isolamento dos fungos. Foram isolados 106 fungos associados às essências florestais, representados por nove gêneros e por dezesseis espécies. Paecilomyces variotii e Lasiodiplodia theobromae foram as espécies fúngicas mais freqüentemente associadas às espécies florestais. Ceiba pentandra e Hura crepitans foram as essências florestais que apresentaram maior diversidade de fungos, com sete espécies cada uma. Hymeneae courbaril foi a que apresentou menor diversidade de fungo, com apenas uma espécie.
In order to know a diversity and incidence of wood mould and wood stain fungi from Amazonian wood, the survey and identification of these kinds of fungi were carried out in twelve wood species from four wood industries of Manaus, Amazonas. Five logs were randomly selected. Wood samples were prepared and transported to the Wood Pathology Laboratory at the Forest Product Research Center of the National Institute for Amazonian Research, where the isolation and identification of the fungi were conducted. One hundred and six fungi associated to the wood species were isolated, represented by nine genera and sixteen fungi species. Paecilomyces variotti and Lasiodiplodia theobromae were the more representative fungi, associated with ten and seven wood species respectively. Ceiba pentandra e Hura crepitans were the wood species with highest diversity of fungi, with seven species each. Hymenae courbaril was the wood that presented the lowest diversity of fungi, with only one fungus.
Subject(s)
Wood , Amazonian Ecosystem , Conservation of Natural Resources , Biodiversity , FungiABSTRACT
As frutas e seus derivados säo em geral alimentos ácidos, sendo contaminados principalmente, por bactérias lácticas, leveduras e fungos filamentosos. Neste trabalho foram isolados e identificados os microorganismos presentes em frutas e polpas de frutas e no ambiente industrial de seu processamento, sendo detectado predomínio de fungos nas amostras analisadas. As bactérias lácticas tiveram maior incidência nas superfícies analisadas