Anti-Acanthamoeba castellanii activity of alkaloid-enriched extracts and lycorine from the Amaryllidaceae species

Abstract Free-living amoebae of the genus Acanthamoeba are the causative agents of granulomatous encephalitis and keratitis, severe human infections. Bioactive compounds from plants are recognized as an alternative source for the development of new drugs. The Amaryllidaceae is a botanical family able to synthesize a very specific and consistent group of biologically active isoquinoline-like alkaloids. The alkaloidal fractions from the Brazilian species Hippeastrum canastrense, H. diniz-cruziae, H. puniceum, and Crinum x amabile, along with the alkaloid lycorine, were investigated against Acanthamoeba castellanii. The in vitro assays were performed with distinct concentrations of lycorine and alkaloidal fractions, while the cell viability was evaluated by the MTT method upon MDCK cells. Chlorhexidine 0.02% was used as the positive control. The effect of alkaloid fractions was concentration dependent, and 2000 µg mL-1 of H. canastrense and H. diniz-cruziae provided a 100% inhibition. At concentrations of 250, 500, and 1000 µg mL-1, the H. diniz-cruziae alkaloidal fraction showed the lowest cytotoxic effect (5%-7%) and remarkable anti-amoebic activity, demonstrating values of IC50 285.61 µg mL-1, low cytotoxicity (5%-7%), and selectivity index (7.0). Taken together, the results are indicative of the great potential that the alkaloids from H. diniz-cruziae have as new candidates for anti-amoebicidal compounds

Antibacterial, antifungal and cytotoxic activities exhibited by endophytic fungi from the Brazilian marine red alga Bostrychia tenella (Ceramiales)

Abstract Marine environment is one of the most important sources regarding natural products research. Besides, marine microorganisms have been denominated as a talented natural source for discovery of new leads. Although the association of macroalgae and fungi has been described regarding ecological issues, there is a lack of studies about marine seaweed endophytic fungi. In this context, the goal of this study was to evaluate cytotoxic, antifungal and antibacterial activities of endophytic fungi isolated from the Brazilian marine seaweed Bostrychia tenella (J.V. Lamouroux) J. Agardh (Ceramiales, Rhodophyta). Forty-five endophytic microorganism strains were isolated from B. tenella. Crude extracts and organic fractions of ten selected strains were obtained after growth in rice medium. Samples were evaluated for cytotoxicity, antifungal and antibacterial assays. Penicillium strains showed positive results in a diversity of assays, and other five strains were active in at least one test. In addition, cytochalasin D was isolated from Xylaria sp. This alga is composed of a microbiological potential, since its endophytic strains exhibited remarkable biological properties. Moreover, cytochalasin D isolation has confirmed chemical potential of marine endophytic strains. This is the first study in which cultured fungi isolates from the Brazilian macroalga B. tenella were evaluated concerning biological properties. Results corroborated that this species could be a pharmaceutical source from marine environment. Furthermore, Acremonium implicatum is being firstly described as marine endophyte and Xylaria sp., Trichoderma atroviride and Nigrospora oryzae as marine seaweed endophytes. Thus, this work reports the first study relating detailed isolation, cultivation and biological evaluation (cytotoxic, antifungal and antibacterial) of endophytes Penicillium decaturense and P. waksmanii from the Brazilian marine red alga B. tenella. We are also reporting the isolation of cytochalasin D, a known antitumor and antibiotic compound, from Xylaria sp. strain. Despite widespread prevalence in terrestrial and marine habitats, this present work describes the first occurrence of cytochalasin D as a metabolite from marine seaweed endophyte.

Endophytic microorganisms from leaves of Spermacoce verticillata (L.): Diversity and antimicrobial activity.

Endophytic microorganisms from the base and top leaves of Spermacoce verticillata were isolated and the antimicrobial potential was evaluated. A total of 56 strains were isolated in pure culture, 44 fungi and 12 actinobacteria. The isolation frequency was higher in the base leaves (12.5%), when compared to the top leaves (3.05%). Among all fungi and actionobacteria identified, the majority belonged to the genus Guignardia (25%) and Microbispora (41.66%), respectively. The antimicrobial screening was firstly evaluated by agar plug assay and showed that 28.57% of the isolates presented activity mainly against gram positive bacteria Staphylococus aureus (ATCC-6538) and Bacillus subtilis (UFPEDA-16). The microorganisms that presented the best activities were then selected, evaluated in different culture media broth, and tested by disk diffusion assay using their fermented broth. The microorganisms selected for this assay exhibited antimicrobial activity mainly for Bacillus subtilis (UFPEDA-16). Since many isolates showed inhibitory activity against pathogenic microorganisms, it is suggestive that endophytic microorganisms from S. verticillata could be an interesting source to explore for bioactive metabolites and new tools need to be employed to explore the real potential of these microorganisms.

Bioactive extracts and chemical constituents of two endophytic strains of Fusarium oxysporum

Ethyl acetate extracts of cultures grown in liquid Czapek and on solid rice media of the fungal endophyte Fusarium oxysporum SS46 isolated from the medicinal plant Smallanthus sonchifolius (Poepp.) H. Rob., Asteraceae, exhibited considerable cytotoxic activity when tested in vitro against human cancer cells. Chromatographic separation yielded anhydrofusarubin (1) and beauvericin (2) that were identified based on their ¹H and 13C NMR data. Compounds 1 and 2 showed the strongest cytotoxic activity against different cancer cell lines. Compound 2 also showed promising activity against Leishmania braziliensis. Hexanic extract of F. oxysporum SS50 grown on solid rice media also afforded a mixture of compounds that displayed cytotoxic activity against different cancer cell lines. Chemical analysis of the mixture of compounds, investigated by gas chromatography-mass spectrometry (GC-MS), showed that there was a predominance of methyl esters of fatty acids and alkanes.

Analysis of 16S rRNA and mxaF genes reveling insights into Methylobacterium niche-specific plant association

The genus Methylobacterium comprises pink-pigmented facultative methylotrophic (PPFM) bacteria, known to be an important plant-associated bacterial group. Species of this group, described as plant-nodulating, have the dual capacity of producing cytokinin and enzymes, such as pectinase and cellulase, involved in systemic resistance induction and nitrogen fixation under specific plant environmental conditions. The aim hereby was to evaluate the phylogenetic distribution of Methylobacterium spp. isolates from different host plants. Thus, a comparative analysis between sequences from structural (16S rRNA) and functional mxaF (which codifies for a subunit of the enzyme methanol dehydrogenase) ubiquitous genes, was undertaken. Notably, some Methylobacterium spp. isolates are generalists through colonizing more than one host plant, whereas others are exclusively found in certain specific plant-species. Congruency between phylogeny and specific host inhabitance was higher in the mxaF gene than in the 16S rRNA, a possible indication of function-based selection in this niche. Therefore, in a first stage, plant colonization by Methylobacterium spp. could represent generalist behavior, possibly related to microbial competition and adaptation to a plant environment. Otherwise, niche-specific colonization is apparently impelled by the host plant.

Avaliação microbiológica e físico-química de méis de jataí (Tetragonisca angustula) e de Apis mellifera do Estado de Mato Grosso do Sul / Microbiological evaluation and physicist-chemistry of honeys of jatai (Tetragonisca angustula) and Apis mellifera of the State of Mato Grosso do Sul

As abelhas jataí produzem mel com propriedades diferentes das abelhas africanas, sendo muito apreciado pelo seu sabor peculiar e pelas propriedades a ele atribuídas, muito embora poucos dados se dispõem sobre as qualidades físico-químicas e biológicas desse mel. Assim, os objetivos do trabalho foram avaliar o grau de contaminação de amostras de méis de abelhas jataí e de Apis mellifera de diferentes regiões do Estado de Mato Grosso do Sul, determinar alguns parâmetros físico-químicos, os quais podem fornecer dados para diferenciar os dois tipos de méis, bem como apresentar alguma correlação com a possível e/ou sobrevivência de microrganismos. Os méis analisados apresentaram pouca contaminação microbiana e não se mostraram contaminados por coliformes fecais ou coliformes a 45ºC, indicando produção e manipulação em boas condições higiênico-sanitárias. Através dos valores médios da umidade, acidez e Brix, os méis de jataí mostraram-se mais aquoso e mais ácidos que os de Apis mellifera e essas variáveis foram consideradas importantes para a diferenciação desses dois tipos de méis.