Leishmanicidal compounds of Nectria pseudotrichia, an endophytic fungus isolated from the plant Caesalpinia echinata (Brazilwood)

BACKGROUND In a screen of extracts from plants and fungi to detect antileishmanial activity, we found that the ethyl acetate extract of the fungus Nectria pseudotrichia, isolated from the tree Caesalpinia echinata (Brazilwood), is a promising source of bioactive compounds. OBJECTIVES The aims of this study were to isolate and determine the chemical structures of the compounds responsible for the antileishmanial activity of the organic extract from N. pseudotrichia. METHODS Compounds were isolated by chromatographic fractionation using semi-preparative high-performance liquid chromatography, and their chemical structures were determined by analytical and spectral data and by comparison with published data. The antileishmanial activity of the isolated compounds was evaluated in intracellular amastigote forms of Leishmania (Viannia) braziliensis expressing firefly luciferase as reporter gene, and cytotoxicity was determined in Vero and THP-1 mammalian cell lines by MTT assay. FINDINGS Fractionation of the extract yielded seven compounds: 10-acetyl trichoderonic acid A (1), 6′-acetoxy-piliformic acid (2), 5′,6′-dehydropiliformic acid (3), piliformic acid (4), hydroheptelidic acid (5), xylaric acid D (6), and cytochalasin D (7). Compounds 1, 2 and 3 are reported here for the first time. Compounds 1, 2, and 5 were more active, with IC50 values of 21.4, 28.3, and 24.8 µM, respectively, and showed low toxicity to Vero and THP-1 cells. MAIN CONCLUSIONS N. pseudotrichia produces secondary metabolites that are more toxic to intracellular amastigote forms of L. (V.) braziliensis than to mammalian cells.

Leishmanicidal compounds of Nectria pseudotrichia, an endophytic fungus isolated from the plant Caesalpinia echinata (Brazilwood).

BACKGROUND In a screen of extracts from plants and fungi to detect antileishmanial activity, we found that the ethyl acetate extract of the fungus Nectria pseudotrichia, isolated from the tree Caesalpinia echinata (Brazilwood), is a promising source of bioactive compounds. OBJECTIVES The aims of this study were to isolate and determine the chemical structures of the compounds responsible for the antileishmanial activity of the organic extract from N. pseudotrichia. METHODS Compounds were isolated by chromatographic fractionation using semi-preparative high-performance liquid chromatography, and their chemical structures were determined by analytical and spectral data and by comparison with published data. The antileishmanial activity of the isolated compounds was evaluated in intracellular amastigote forms of Leishmania (Viannia) braziliensis expressing firefly luciferase as reporter gene, and cytotoxicity was determined in Vero and THP-1 mammalian cell lines by MTT assay. FINDINGS Fractionation of the extract yielded seven compounds: 10-acetyl trichoderonic acid A (1), 6'-acetoxy-piliformic acid (2), 5',6'-dehydropiliformic acid (3), piliformic acid (4), hydroheptelidic acid (5), xylaric acid D (6), and cytochalasin D (7). Compounds 1, 2 and 3 are reported here for the first time. Compounds 1, 2, and 5 were more active, with IC50 values of 21.4, 28.3, and 24.8 µM, respectively, and showed low toxicity to Vero and THP-1 cells. MAIN CONCLUSIONS N. pseudotrichia produces secondary metabolites that are more toxic to intracellular amastigote forms of L. (V.) braziliensis than to mammalian cells.

Antimicrobial potential of actinobacteria isolated from the rhizosphere of the Caatinga biome plant Caesalpinia pyramidalis Tul.

Actinobacteria are known to produce various secondary metabolites having antibiotic effects. This study assessed the antimicrobial potential of actinobacteria isolated from the rhizosphere of Caesalpinia pyramidalis Tul. from the Caatinga biome. Sixty-eight actinobacteria isolates were evaluated for antimicrobial activity against different microorganisms by disk diffusion and submerged fermentation, using different culture media, followed by determination of minimum inhibitory concentration (MIC) and chemical prospecting of the crude extract. Of the isolates studied, 52.9% of those isolated at 37°C and 47.05% of those isolated at 45°C had activity against Bacillus subtilis, Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), Fusarium moniliforme, and Candida albicans. When compared with others actinobacteria, the isolate C1.129 stood out with better activity and was identified by 16S rDNA gene analysis as Streptomyces parvulus. The crude ethanol extract showed an MIC of 0.97 µg/mL for MRSA and B. subtilis, while the ethyl acetate extract showed MIC of 3.9 µg/mL for S. aureus and MRSA, showing the greatest potential among the metabolites produced. Chemical prospecting revealed the presence of mono/sesquiterpenes, proanthocyanidin, triterpenes, and steroids in both crude extracts. This study evaluates S. parvulus activity against multi-resistant microorganisms such as MRSA. Thus, it proves that low-fertility soil, as is found in the Caatinga, may contain important microorganisms for the development of new antimicrobial drugs.

Bioactive endophytic fungi isolated from Caesalpinia echinata Lam. (Brazilwood) and identification of beauvericin as a trypanocidal metabolite from Fusarium sp.

Aiming to identify new sources of bioactive secondary metabolites, we isolated 82 endophytic fungi from stems and barks of the native Brazilian tree Caesalpinia echinata Lam. (Fabaceae). We tested their ethyl acetate extracts in several in vitro assays. The organic extracts from three isolates showed antibacterial activity against Staphylococcus aureus and Escherichia coli [minimal inhibitory concentration (MIC) 32-64 µg/mL]. One isolate inhibited the growth of Salmonella typhimurium (MIC 64 µg/mL) and two isolates inhibited the growth of Klebsiella oxytoca (MIC 64 µg/mL), Candida albicans and Candida tropicalis (MIC 64-128 µg/mL). Fourteen extracts at a concentration of 20 µg/mL showed antitumour activities against human breast cancer and human renal cancer cells, while two isolates showed anti-tumour activities against human melanoma cancer cells. Six extracts were able to reduce the proliferation of human peripheral blood mononuclear cells, indicating some degree of selective toxicity. Four isolates were able to inhibit Leishmania (Leishmania) amazonensis and one isolate inhibited Trypanosoma cruzi by at least 40% at 20 µg/mL. The trypanocidal extract obtained from Fusarium sp. [KF611679] culture was subjected to bioguided fractionation, which revealed beauvericin as the compound responsible for the observed toxicity of Fusarium sp. to T. cruzi. This depsipeptide showed a half maximal inhibitory concentration of 1.9 µg/mL (2.43 µM) in a T. cruzi cellular culture assay.

Antioxidant and antimicrobial properties of ethanolic extract of Libidibia ferrea pods / Propriedade antioxidante e antimicrobiana do extrato etanólico de vagens de Libidibia ferrea

Libidibia ferrea is a typical plant of northern and north-east Brazil and has well-known medicinal properties that can be found in ethanolic extracts such as antipyretic, anti-inflammatory and anti-rheumatic compounds. This study seeks to evaluate the potential antioxidant and antimicrobial activity of ethanol extracts from Libidibia ferrea pods. The pods were used for the preparation of ethanolic extracts of L. ferrea which is used to determine biological activity, and measure their total phenolic content. For quantification of antioxidant methods of DPPH, ABTS and ß-carotene were employed and the method of minimum inhibitory concentration (MIC) to measure the antimicrobial activity. The ethanolic extract of L. ferrea showed excellent antioxidant activity, as well as other similar substances commonly used for this purpose such as citric acid and trolox. In addition, it had antimicrobial activity against Gram-negative and Gram-positive bacteria.(AU)

Libidibia ferrea é uma planta típica do norte e nordeste do Brasil conhecida por suas propriedades medicinais que podem ser encontrados em extratos alcoólicos, possui compostos antipiréticos, anti-inflamatórios e antirreumáticos. Este estudo visa avaliar o potencial antioxidante e atividade antimicrobiana do extrato etanólico de vagens de Libidibia ferrea. As vagens foram empregadas na preparação de extrato etanólico de L. férrea, utilizado na determinação da atividade biológica, e para mensurar seu conteúdo fenólico total. Os métodos DPPH, ABTS e ß-caroteno foram utilizados para quantificar a atividade antioxidante, e o método da Concentração Inibitória Mínima (CIM) para avaliar a atividade antimicrobiana. O extrato etanólico de L. ferrea apresentou atividade antioxidante excelente, bem como outras substâncias compatíveis normalmente utilizadas para este fim, tais como ácido cítrico e trolox. Além disso, demonstrou atividade antimicrobiana contra bactérias Gram-negativas e Gram-positivas.(AU)

Molecular characterization of Cryptococcus gattii genotype AFLP6/VGII isolated from woody debris of divi-divi (Caesalpinia coriaria), Bonaire, Dutch Caribbean / Caracterización molecular del genotipo AFLP6/VGII Cryptococcus gattii aislado de restos de madera de divi-divi (Caesalpinia coriaria), Bonaire, Caribe Holandés

Background. The basidiomycetous yeast Cryptococcus gattii is an emerging and primary pathogen. There is a lack of information about its environmental spread outside outbreak regions in Mediterranean Europe, North and South America. Environmental sampling for C. gattii and molecular characterization of the obtained isolates will provide an insight into the global spread of the various genotypes. Methods. Woody debris of native divi-divi (Caesalpinia coriaria) trees were sampled across Bonaire, Dutch Caribbean. Colonies suspected for Cryptococcus species were subjected to standard mycology investigations and identification by matrix-assisted laser desorption ionization-time of flight mass spectrometry. Isolates identified as C. gattii were subjected to amplified fragment length polymorphism genotyping, mating-type analysis and multi-locus sequence typing. Results. Ten colonies of C. gattii were cultured from different trunk hollows of the same divi-divi tree. Molecular characterization showed that all isolates were genotype AFLP6/VGII and mating-type α. Multi-locus sequence typing revealed that all isolates were genetically indistinguishable from each other. Conclusions. C. gattii is present in the environment of Bonaire, which suggests that this yeast is likely to be present in the environment of other Caribbean islands (AU)

Antecedentes. La levadura Cryptococcus gattii es un basidiomiceto emergente y patógeno primario. Existe poca información acerca de su dispersión en medio ambiente fuera de las regiones con brotes descritos por esta levadura en la Europa mediterránea, Norte y Sur de América. Los muestreos del medio ambiente para la búsqueda de C. gattii y la caracterización molecular de los aislamientos obtenidos puede proveer de una visión global sobre la dispersión de varios de sus genotipos. Métodos. Se tomaron muestras de residuos de madera de árboles nativos divi-divi (Caesalpinia coriaria) en Bonaire, Caribe Holandés. Las colonias susceptibles de pertenecer a las especies de Cryptococcus se sometieron a un estudio micológico estándar e identificación por espectrometría de masas MALDI-TOF (Matrix-Assisted Laser Desorption Ionization-Time of Flight). Los aislamientos identificados como C. gattii se sometieron a genotipado mediante AFLP (Amplified Fragment Length Polymorphism), obtención del tipo sexual y MLST (Multi-locus Sequence Typing). Resultados. Se obtuvieron diez colonias de C. gattii en el cultivo de nuestras de diferentes agujeros de un mismo árbol divi-divi. La caracterización molecular mostró que todos los aislamientos eran genotipo AFLP6/VGII y tipo sexual α. El tipado mediante MLST reveló que todos los aislamientos eran genéticamente indistinguibles unos de otros. Conclusiones. C. gattii está presente en el medio ambiente de Bonaire, lo que sugiere que esta levadura podría estar presente en el ambiente de otras islas del Caribe (AU)

Molecular characterization of Cryptococcus gattii genotype AFLP6/VGII isolated from woody debris of divi-divi (Caesalpinia coriaria), Bonaire, Dutch Caribbean.

BACKGROUND: The basidiomycetous yeast Cryptococcus gattii is an emerging and primary pathogen. There is a lack of information about its environmental spread outside outbreak regions in Mediterranean Europe, North and South America. Environmental sampling for C. gattii and molecular characterization of the obtained isolates will provide an insight into the global spread of the various genotypes. METHODS: Woody debris of native divi-divi (Caesalpinia coriaria) trees were sampled across Bonaire, Dutch Caribbean. Colonies suspected for Cryptococcus species were subjected to standard mycology investigations and identification by matrix-assisted laser desorption ionization-time of flight mass spectrometry. Isolates identified as C. gattii were subjected to amplified fragment length polymorphism genotyping, mating-type analysis and multi-locus sequence typing. RESULTS: Ten colonies of C. gattii were cultured from different trunk hollows of the same divi-divi tree. Molecular characterization showed that all isolates were genotype AFLP6/VGII and mating-type α. Multi-locus sequence typing revealed that all isolates were genetically indistinguishable from each other. CONCLUSIONS: C. gattii is present in the environment of Bonaire, which suggests that this yeast is likely to be present in the environment of other Caribbean islands.