RESUMO
Polysaccharides from P. eryngii mushroom were selectively extracted using low-cost technologies (water at different conditions of temperature and pressure). Mannogalactan was the main polysaccharide in cold-water extracted fraction (CWEF), while a linear (1â6)-ß-d-glucan was the main polymer in hot-water extracted fraction (HWEF). Autoclave-extracted fraction (AEF) contained a mixture of at least four different α- and ß-glucans. The report of linear (1â6)-ß-glucan and linear (1â3)-ß-glucan is a new finding for P. eryngii fruiting bodies. The immunostimulatory properties of the fractions on THP-1 macrophages were studied. All fractions at 50, 250 and 500 µg/mL were not cytotoxic and produced different stimulus on NO, IL-1ß and IL-10 secretion by the cells. Thus, our results showed that it is possible to concentrate different P. eryngii polysaccharides in selected fractions using a simple and low-cost procedure. Since biological effects depends on the polysaccharide structure, this technique allows the obtainment of fractions with distinct immunomodulatory activities.
Assuntos
Carpóforos/química , Polissacarídeos Fúngicos/farmacologia , Fatores Imunológicos , Macrófagos/efeitos dos fármacos , Pleurotus/química , Polissacarídeos , beta-Glucanas , Misturas Complexas/química , Misturas Complexas/isolamento & purificação , Misturas Complexas/farmacologia , Polissacarídeos Fúngicos/isolamento & purificação , Humanos , Fatores Imunológicos/isolamento & purificação , Fatores Imunológicos/farmacologia , Imunomodulação , Estrutura Molecular , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Relação Estrutura-Atividade , Células THP-1 , beta-Glucanas/isolamento & purificação , beta-Glucanas/farmacologiaRESUMO
The cyanobacterium Radiocystis fernandoi has been frequently identified in cyanobacterial blooms in Brazil. Recently, R. fernandoi strain R28, which produces microcystin (MC)-RR and MC-YR, was isolated from the Furnas reservoir, Minas Gerais, Brazil. The present study evaluated the hematological variables and erythrocyte antioxidant responses, lipid peroxidation (LPO), and genotoxicity in a neotropical fish (Hoplias malabaricus) after acute and subchronic exposure to a crude extract (CE) of R. fernandoi strain R28. Acute exposure (12 or 96â¯h) consisted of a single intraperitoneal (i.p.) CE injection, and subchronic exposure consisted of one i.p. CE injection every 72â¯h for 30 days. After acute exposure, fish exhibited macrocytic anemia (12â¯h post-injection) followed by normocytic anemia (96â¯h post-injection). The increased activity of superoxide dismutase, glutathione peroxidase, glutathione-S-transferase, and the glutathione level in the erythrocytes did not prevent oxidative stress, manifested as lipid peroxidation and elevated DNA damage after acute exposure. After subchronic exposure, the hematological variables recovered, and the absence of erythrocyte oxidative stress suggests possible modulation by other biological factors, including a possible decrease in MC uptake by the cells and/or increasing detoxification efficiency that precludes erythrocyte damage.
Assuntos
Misturas Complexas/toxicidade , Cianobactérias/química , Eritrócitos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Animais , Misturas Complexas/química , Misturas Complexas/isolamento & purificação , Dano ao DNA/efeitos dos fármacos , Peixes , Peroxidação de Lipídeos/efeitos dos fármacos , Testes de ToxicidadeRESUMO
Tropical ecosystems hold an extremely diverse array of endophytic fungi, but their potential use still remains to be explored. In this study, we isolated an endophytic fungus from the leaves of Otoba gracilipes, a medicinal tree from a tropical rainforest in Colombia. Following isolation and cultivation, we evaluated its extracellular crude extract for antioxidant activity. Using traditional and molecular methods (ITS1, NL1 regions), the endophyte was identified as Fusarium oxysporum. Fresh spores from the fungal isolate were inoculated in liquid media (potato dextrose broth [PDB] and potato dextrose-yeast extract broth [PDYB]) and centrifuged for recovering extracellular polysaccharides from the exhausted medium after 30 days of cultivation. Crude extracts were recovered, purified, lyophilized, and evaluated for their ability to inactivate the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH). The extracts obtained from PDB culture media had a 51.5% of scavenging effect on DPPH after 5 min of reaction compared with the extracts from PDBY (26.4%), which suggests a high antioxidant potential of these fungal extracts. Thus, our results suggest other fungi from tropical ecosystems should be explored as potential sources of novel enzymes and other metabolites with bioactivity.
Assuntos
Antioxidantes/metabolismo , Produtos Biológicos/metabolismo , Endófitos/metabolismo , Fusarium/metabolismo , Myristicaceae/microbiologia , Antioxidantes/isolamento & purificação , Produtos Biológicos/isolamento & purificação , Colômbia , Misturas Complexas/isolamento & purificação , Misturas Complexas/metabolismo , Meios de Cultura/química , Endófitos/classificação , Endófitos/genética , Endófitos/isolamento & purificação , Fusarium/classificação , Fusarium/genética , Fusarium/isolamento & purificação , Metagenômica , Técnicas Microbiológicas , Folhas de Planta/microbiologia , Clima TropicalRESUMO
Vibrio diabolicus A1SM3 strain was isolated from a sediment sample from Manaure Solar Saltern in La Guajira and the produced crude extracts have shown antibacterial activity against methicillin-resistant Staphylococcus aureus and cytotoxic activity against human lung cell line. Thus, the aim of this research was to identify the main compound responsible for the biological activity observed and to systematically study how each carbon and nitrogen source in the growth media, and variation of the salinity, affect its production. For the characterization of the bioactive metabolites, 15 fractions obtained from Vibrio diabolicus A1SM3 crude extract were analyzed by HPLC-MS/MS and their activity was established. The bioactive fractions were dereplicated with Antibase and Marinlit databases, which combined with nuclear magnetic resonance (NMR) spectra and fragmentation by MS/MS, led to the identification of 2,2-di(3-indolyl)-3-indolone (isotrisindoline), an indole-derivative antibiotic, previously isolated from marine organisms. The influence of the variations of the culture media in isotrisindoline production was established by molecular network and MZmine showing that the media containing starch and peptone at 7% NaCl was the best culture media to produce it. Also, polyhydroxybutyrates (PHB) identification was established by MS/MS mainly in casamino acids media, contributing to the first report on PHB production by this strain.
Assuntos
Antibacterianos/biossíntese , Antibacterianos/farmacologia , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Técnicas Bacteriológicas/métodos , Vibrio/química , Vibrio/metabolismo , Alcaloides/biossíntese , Alcaloides/isolamento & purificação , Alcaloides/farmacologia , Antibacterianos/isolamento & purificação , Antineoplásicos/isolamento & purificação , Organismos Aquáticos/microbiologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Misturas Complexas/química , Misturas Complexas/isolamento & purificação , Meios de Cultura , Humanos , Hidroxibutiratos/química , Hidroxibutiratos/farmacologia , Isoindóis/isolamento & purificação , Isoindóis/metabolismo , Modelos Moleculares , Poliésteres/química , Poliésteres/farmacologia , Poli-Hidroxialcanoatos/química , Poli-Hidroxialcanoatos/farmacologia , Proibitinas , SalinidadeRESUMO
Phenolic (free, conjugated and bound) and carotenoid extracts from microalgae Nannochloropsis sp. and Spirulina sp. were investigated regarding their potential to mitigate contamination by Fusarium complex fungal pathogens. Free phenolic acid extracts from both microalgae were the most efficient, promoting the lowest mycelial growth rates of 0.51 cm day- 1 (Spirulina sp.) and 0.78 cm day- 1 (Nannochloropsis sp.). An experiment involving natural free phenolic acid extracts and synthetic solutions was carried out based on the natural phenolic acid profile. The results revealed that the synthetic mixtures of phenolic acids from both microalgae were less efficient than the natural extracts at inhibiting fungal growth, indicating that no purification is required. The half-maximal effective concentration (EC50) values of 49.6 µg mL- 1 and 33.9 µg mL- 1 were determined for the Nannochloropsis and Spirulina phenolic acid extracts, respectively. The use of phenolic extracts represents a new perspective regarding the application of compounds produced by marine biotechnology to prevent Fusarium species contamination.
Assuntos
Antifúngicos/isolamento & purificação , Antifúngicos/farmacologia , Fusarium/efeitos dos fármacos , Fusarium/crescimento & desenvolvimento , Spirulina/química , Estramenópilas/química , Misturas Complexas/isolamento & purificação , Misturas Complexas/farmacologia , Micélio/efeitos dos fármacos , Micélio/crescimento & desenvolvimentoRESUMO
Conventional and commercial methods for isolation of nucleic acids are available for fungal samples including entomopathogenic fungi (EPF). However, there is not a unique optimal method for all organisms. The cell wall structure and the wide range of secondary metabolites of EPF can broadly interfere with the efficiency of the DNA extraction protocol. This study compares three commercial protocols: DNeasy® Plant Mini Kit (Qiagen), Wizard® Genomic DNA Purification Kit (Promega), and Axygen™ Multisource Genomic DNA Miniprep Kit (Axygen) and three conventional methods based on different buffers: SDS, CTAB/PVPP, and CTAB/ß-mercaptoethanol versus three cell lysis procedures: liquid nitrogen homogenization and two bead-beating materials (i.e., tungsten-carbide and stainless-steel) for four representative species of EPF (i.e., Beauveria bassiana, Hirsutella citriformis, Isaria javanica, and Metarhizium anisopliae). Liquid nitrogen homogenization combined with DNeasy® Plant Mini Kit (i.e., QN) or SDS buffer (i.e., SN) significantly improved the yield with a good purity (~1.8) and high integrity (>20,000â¯bp) of genomic DNA in contrast with other methods, also, these results were better when compared with the two bead-beating materials. The purified DNA was evaluated by PCR-based techniques: amplification of translation elongation factor 1-α (TEF) and two highly sensitive molecular markers (i.e., ISSR and AFLP) with reliable and reproducible results. Despite a variation in yield, purity, and integrity of extracted DNA across the four species of EPF with the different DNA extraction methods, the SN and QN protocols maintained a high-quality of DNA which is required for downstream molecular applications.
Assuntos
DNA Fúngico/isolamento & purificação , Fungos/genética , Genômica/métodos , Misturas Complexas/isolamento & purificação , Reação em Cadeia da PolimeraseRESUMO
This study evaluated the use of commercial rhamnolipid biosurfactant supplementation in the phytoremediation of a soil via sunflower (Helianthus annuus L.) cultivation. The soil, obtained from an industrial area, was co-contaminated with heavy metals and petroleum hydrocarbons. The remediation tests were monitored for 90 days. The best results for removal of contaminants were obtained from the tests in which the sunflower plants were cultivated in soil with 4 mg kg-1 of the rhamnolipid. Under these conditions, reductions of 58% and 48% were obtained in the total petroleum hydrocarbon (TPH) and polycyclic aromatic hydrocarbon (PAH) concentrations, respectively; reductions in the concentrations of the following metals were also achieved: Ni (41%), Cr (30%), Pb (29%), and Zn (20%). The PCR-DGGE analysis of soil samples collected before and after the treatments verified that the plant cultivation and biosurfactants supplementation had little effect on the structure of the dominant bacterial community in the soil. The results indicated that sunflower cultivation with the addition of a biosurfactant is a viable and efficient technology to treat soils co-contaminated with heavy metals and petroleum hydrocarbons.
Assuntos
Misturas Complexas/isolamento & purificação , Helianthus/química , Resíduos Industriais , Indústria de Petróleo e Gás , Poluentes do Solo/isolamento & purificação , Solo/química , Tensoativos/química , Biodegradação Ambiental , Misturas Complexas/química , Humanos , Hidrocarbonetos/análise , Hidrocarbonetos/isolamento & purificação , Metais Pesados/análise , Metais Pesados/isolamento & purificação , Petróleo/análise , Hidrocarbonetos Policíclicos Aromáticos/análise , Hidrocarbonetos Policíclicos Aromáticos/isolamento & purificação , Poluentes do Solo/análiseRESUMO
Increased resistance to the first-line treatment against P. falciparum malaria, artemisinin-based combination therapies, has been reported. Here, we tested the effect of crude ethanolic extract of the fungus Trichoderma stromaticum (Ext-Ts) on the growth of P. falciparum NF54 in infected human red blood cells (ihRBCs) and its anti-malarial and anti-inflammatory properties in a mouse model of experimental cerebral malaria. For this purpose, ihRBCs were treated with Ext-Ts and analysed for parasitaemia; C57BL/6 mice were infected with P. berghei ANKA (PbA), treated daily with Ext-Ts, and clinical, biochemical, histological and immunological features of the disease were monitored. It was observed that Ext-Ts presented a dose-dependent ability to control P. falciparum in ihRBCs. In addition, it was demonstrated that Ext-Ts treatment of PbA-infected mice was able to increase survival, prevent neurological signs and decrease parasitaemia at the beginning of infection. These effects were associated with systemically decreased levels of lipids and IFN-γ, ICAM-1, VCAM-1 and CCR5 cerebral expression, preserving blood brain barrier integrity and attenuating the inflammatory lesions in the brain, liver and lungs. These results suggest that Ext-Ts could be a source of immunomodulatory and antimalarial compounds that could improve the treatment of cerebral malaria.
Assuntos
Anti-Inflamatórios/farmacologia , Antimaláricos/farmacologia , Misturas Complexas/farmacologia , Malária Cerebral/tratamento farmacológico , Trichoderma/química , Animais , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/isolamento & purificação , Antimaláricos/administração & dosagem , Antimaláricos/isolamento & purificação , Encéfalo/parasitologia , Encéfalo/patologia , Misturas Complexas/administração & dosagem , Misturas Complexas/isolamento & purificação , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Eritrócitos/parasitologia , Histocitoquímica , Humanos , Imuno-Histoquímica , Malária Cerebral/parasitologia , Malária Cerebral/patologia , Camundongos Endogâmicos C57BL , Parasitemia/tratamento farmacológico , Plasmodium falciparum/efeitos dos fármacos , Análise de Sobrevida , Resultado do TratamentoRESUMO
Despite the great advances in chemotherapeutics, infectious diseases are still one of the leading causes of death worldwide. Among some of the clinically relevant pathogens, methicillin-resistant Staphylococcus aureus (MRSA) ranks as one of the most difficult bacteria to treat. It is a common cause of skin, soft-tissue, and endovascular infections, as well as pneumonia, septic arthritis, endocarditis, osteomyelitis, and sepsis. The research on Basidiomycota is extensive; many species show a broad spectrum of pharmacological activities, including antimicrobial activity. The vast majority of the literature to date generally focuses on screening the antibacterial properties of mushroom extracts. A gap still exists in the identification of the individual compounds responsible for these properties, and few low molecular weight compounds have been described. Gymnopilus junonius, the big laughter mushroom, grows wild in Uruguay, especially on Eucalyptus spp. plantations; it is known as the "eucalyptus fungus." In this work, we report the bioguided isolation, structural elucidation, and antistaphylococcal activity of the main antimicrobial components of fresh basidiocarps of G. junonius.
Assuntos
Agaricales/química , Antibacterianos/farmacologia , Misturas Complexas/farmacologia , Carpóforos/química , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Agaricales/crescimento & desenvolvimento , Antibacterianos/isolamento & purificação , Misturas Complexas/isolamento & purificação , Eucalyptus/microbiologia , Testes de Sensibilidade Microbiana , UruguaiRESUMO
Piptoporus betulinus has been used in folk medicine for millennia. However, no data currently exist regarding its potential cardiovascular activity. In this work, the crude ethanolic extract and fractions (hexane, ethyl acetate, and water) with increased polarity from the partitioning process, as well as stigmasterol (the major metabolite isolated from P. betulinus), were administered orally at different doses to normotensive male Wistar rats an hour before recording mean arterial pressure, heart rate, renal blood flow, renal vascular conductance, arterial blood flow, and arterial vascular conductance. The acute oral administration of crude ethanolic extract and all fractions did not alter mean arterial pressure when compared with the control group, which received a vehicle. In addition, subchronic (14 days) oral administration of crude ethanolic extract, fractions, and stigmasterol did not alter cardiovascular parameters. In conclusion, our findings demonstrate that oral administration of organic extracts of P. betulinus did not induce cardiovascular alterations.
Assuntos
Sistema Cardiovascular/efeitos dos fármacos , Misturas Complexas/administração & dosagem , Polyporales/química , Estigmasterol/administração & dosagem , Administração Oral , Animais , Misturas Complexas/isolamento & purificação , Masculino , Ratos Wistar , Estigmasterol/isolamento & purificaçãoRESUMO
In this work, a saline aqueous solution of phenol, 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4-dichlorophenol (2,4-DCP) was treated by the photo-Fenton process in a falling-film solar reactor. The influence of the parameters such as initial pH (5-7), initial concentration of Fe2+ (1-2.5mM) and rate of H202 addition (1.87-3.74mmol min-1) was investigated. The efficiency of photodegradation was determined from the removal of dissolved organic carbon (DOC), described by the species degradation of phenol, 2,4-D and 2,4-DCP. Response surface methodology was employed to assess the effects of the variables investigated, i.e. [Fe2+], [H202] and pH, in the photo-Fenton process with solar irradiation. The results reveal that the variables' initial concentration of Fe2+ and H202 presents predominant effect on pollutants' degradation in terms of DOC removal, while pH showed no influence. Under the most adequate experimental conditions, about 85% DOC removal was obtained in 180 min by using a reaction system employed here, and total removal of phenol, 2,4- and 2,4-DCP mixture in about 30min.
Assuntos
Ácido 2,4-Diclorofenoxiacético/química , Peróxido de Hidrogênio/química , Ferro/química , Fenóis/química , Fotoquímica/instrumentação , Cloreto de Sódio/química , Energia Solar , Purificação da Água/instrumentação , Ácido 2,4-Diclorofenoxiacético/isolamento & purificação , Ácido 2,4-Diclorofenoxiacético/efeitos da radiação , Misturas Complexas/química , Misturas Complexas/isolamento & purificação , Misturas Complexas/efeitos da radiação , Fontes de Energia Elétrica , Desenho de Equipamento , Análise de Falha de Equipamento , Peróxido de Hidrogênio/efeitos da radiação , Ferro/efeitos da radiação , Fenóis/isolamento & purificação , Fenóis/efeitos da radiação , Fotoquímica/métodos , Cloreto de Sódio/efeitos da radiaçãoRESUMO
Oil palm (Elaeis guineensis) fruit was treated with enzymes to facilitate simultaneous recovery of oil and bioactive compounds. Tannase from Paecilomyces variotii, cellulase and pectinase were evaluated for their influence on oil recovery and antioxidant capacity (DPPH), oxidative stability (Rancimat), fatty acid profile, total phenols, total carotenoids and tocols of the oil. Maximum oil recovery (90-93% total oil) was obtained with central composite design using 4% of enzyme preparation (w/w) as 80 U of tannase, 240 U of cellulase and 178 U of pectinase, pH 4, ratio of solution to pulp of 2:1 and 30 min of incubation at 50 °C. Tannase improved the phenolic compounds extraction by 51% and pectinase plus cellulase improved carotene extraction by 153%. Samples treated with tannase showed a 27% and 53% higher antioxidant capacity for the lipophilic and hydrophilic fractions.
Assuntos
Antioxidantes/isolamento & purificação , Araceae/química , Hidrolases de Éster Carboxílico/química , Celulases/química , Frutas/química , Óleos de Plantas/isolamento & purificação , Poligalacturonase/química , Misturas Complexas/química , Misturas Complexas/isolamento & purificação , Óleo de Palmeira , Água/químicaRESUMO
BACKGROUND: Phycobiliproteins are coloured proteins produced by cyanobacteria, which have several applications because of their colour properties. However, there is no available information about the colour stability of phycobiliproteins from Nostoc sp. in food systems. The aim of this work was to study the colour stability of a purple-coloured phycobiliprotein-rich extract from the cyanobacterium Nostoc PCC9205 in acidic solutions and yogurt. RESULTS: Variations of pH for Nostoc PCC9205 extract have shown stability for the L* (lightness) and a* (redness) indexes in the range 1.0-7.0. The b* index (blueness), however, increased at pH values below 4.0, indicating loss of the blue colour. The Nostoc PCC9205 extract was used as colorant in yogurt (pH 4.17) stored for 60 days. Instrumental colour analysis showed no changes for the L* and a* indexes during storage, whereas the b* index changed after 20 days of storage. A multiple comparison test showed colour instability after 20 days of storage. A hedonic scale test performed on the 60th day of storage showed acceptability of the product. CONCLUSIONS: The red component of the phycobiliprotein-rich extract from Nostoc PCC9205 presented an improved stability in acidic media and yogurt compared with the blue component of this extract.
Assuntos
Proteínas de Bactérias/análise , Misturas Complexas/química , Corantes de Alimentos/análise , Nostoc/química , Ficobiliproteínas/análise , Pigmentos Biológicos/análise , Iogurte/análise , Proteínas de Bactérias/biossíntese , Proteínas de Bactérias/química , Proteínas de Bactérias/isolamento & purificação , Brasil , Cor , Misturas Complexas/isolamento & purificação , Corantes de Alimentos/química , Corantes de Alimentos/isolamento & purificação , Corantes de Alimentos/metabolismo , Preferências Alimentares , Qualidade dos Alimentos , Armazenamento de Alimentos , Humanos , Concentração de Íons de Hidrogênio , Nostoc/metabolismo , Ficobiliproteínas/biossíntese , Ficobiliproteínas/química , Ficobiliproteínas/isolamento & purificação , Pigmentos Biológicos/biossíntese , Pigmentos Biológicos/química , Pigmentos Biológicos/isolamento & purificação , Estabilidade Proteica , Sensação , Solubilidade , EspectrofotometriaRESUMO
This study investigated the effects of Agaricus brasiliensis S. Wasser et al. (=Agaricus blazei Murrill sensu Heinem.) aqueous extract on small intestinal sIgA levels, serum TNF-alpha, IFN-gamma and IL-10 levels, splenic index, bacterial translocation, and histology of small intestine, spleen, and liver from mice orally challenged with 10(6) CFU of Salmonella enterica serovar Typhimurium (SEST). Splenic index values as well as sIgA, TNF-alpha, IFN-gamma, and IL-10 levels were not affected by either A. brasiliensis aqueous extract treatment or by pathogenic challenge. Typical colonies of SEST were recovered from liver, spleen, and mesenteric lymph nodes of challenged animals, but there was no significant difference in this translocation between groups treated or not with A. brasiliensis aqueous extract. Translocation was confirmed by histopathological analysis in mice challenged with SEST, which showed small and diffuse foci of mixed inflammatory infiltrate in hepatic parenchyma. In conclusion, A. brasiliensis aqueous extract as tested in the present study did not influence any of the variables selected to evaluate in vivo its immunomodulatory effect suggested in the literature.
Assuntos
Agaricus/química , Translocação Bacteriana , Misturas Complexas/farmacologia , Citocinas/sangue , Salmonelose Animal/imunologia , Salmonella typhimurium/fisiologia , Animais , Peso Corporal/efeitos dos fármacos , Misturas Complexas/isolamento & purificação , Citocinas/efeitos dos fármacos , Citocinas/metabolismo , Imunoglobulina A Secretora/efeitos dos fármacos , Imunoglobulina A Secretora/metabolismo , Interferon gama/sangue , Interferon gama/efeitos dos fármacos , Interleucina-10/sangue , Intestino Delgado/efeitos dos fármacos , Intestino Delgado/imunologia , Fígado/efeitos dos fármacos , Fígado/patologia , Linfonodos/efeitos dos fármacos , Linfonodos/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Modelos Animais , Salmonelose Animal/microbiologia , Salmonella typhimurium/efeitos dos fármacos , Baço/efeitos dos fármacos , Baço/patologia , Fator de Necrose Tumoral alfa/sangue , Fator de Necrose Tumoral alfa/efeitos dos fármacosRESUMO
We investigated in vitro antioxidant activities of 49 endophytic fungi isolated from the liverwort Scapania verrucosa. Based on morphological and molecular identification, the endophytic fungi isolated were classified into seven genera (Hypocrea, Penicillium, Tolypocladium, Chaetomium, Xylaria, Nemania, and Creosphaeria), all belonging to one family (Xylariaceae). By screening with the 2,2'-azino-di(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) decolorization assay, the ethyl acetate extracts of five endophytic fungi (T7, T21, T24, T32, and T38 strains), which exhibited remarkable Trolox equivalent (TE) antioxidant capacity (ranging from 997.06 to 1248.10 µmol TE/g extract), were selected and their antioxidant capacity was further evaluated by assays for 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, hydroxyl radical scavenging, reducing power, and ferrous ion chelating. The ethyl acetate extracts of two endophytic fungi (T24 and T38) were found to have comparable scavenging abilities on both DPPH-free radicals (93.9 and 88.7%, respectively, at 50 µg/mL) and hydroxyl radicals (97.1 and 89.4%, respectively, at 2 mg/mL) when compared with those of the positive controls (ascorbic acid and butylated hydroxytoluene, respectively). Although their reducing powers were similar to that of butylated hydroxytoluene, as indicated by absorbance (0.35 and 0.30 at 50 µg/mL, respectively), only the T38 strain's ethyl acetate extract showed ferrous ion chelating ability (92.9% at 1 mg/mL) comparable to that of the EDTA-2Na control. These endophytic fungi in S. verrucosa are a potential novel source of natural antioxidants.
Assuntos
Antioxidantes/farmacologia , Endófitos/química , Sequestradores de Radicais Livres/farmacologia , Fungos/química , Genes Fúngicos , Hepatófitas/microbiologia , Acetatos , Antioxidantes/isolamento & purificação , Benzotiazóis/química , Benzotiazóis/metabolismo , Compostos de Bifenilo/antagonistas & inibidores , China , Cromanos/farmacologia , Misturas Complexas/isolamento & purificação , Misturas Complexas/farmacologia , Endófitos/classificação , Endófitos/isolamento & purificação , Sequestradores de Radicais Livres/isolamento & purificação , Radicais Livres/antagonistas & inibidores , Fungos/classificação , Fungos/isolamento & purificação , Genes de RNAr , Radical Hidroxila/antagonistas & inibidores , Quelantes de Ferro/isolamento & purificação , Quelantes de Ferro/farmacologia , Oxirredução , Picratos/antagonistas & inibidores , Solventes , Ácidos Sulfônicos/química , Ácidos Sulfônicos/metabolismoRESUMO
Introducción: los moluscos marinos constituyen un reservorio natural de moléculas con potencialidades terapéuticas para el tratamiento de enfermedades infecciosas en momentos en que se han descrito numerosas cepas resistentes a los antibióticos convencionales. Objetivo: comparar 3 soluciones: ácido acético 30 por ciento, metanol 50 por ciento y salina-ácida (NaCl 0,6 mol/L, HCl 1 por ciento) atendiendo a sus capacidades extractivas de moléculas con actividad antibacteriana del molusco marino Cenchritis muricatus. Métodos: para el procesamiento del material biológico se utilizaron las 3 soluciones de extracción y se analizaron los extractos obtenidos de acuerdo con la concentración de proteínas totales y la inhibición del crecimiento bacteriano de cepas de Staphylococcus aureus y Escherichia coli, mediante un bioensayo turbidimétrico en microplacas de 96 pocillos en medio Luria-Bertani. Resultados: se obtuvo mayor concentración de proteínas totales (7,8 mg/mL) con el extracto total de C. muricatus obtenido con la solución salina-ácida. Además con 200 mg/mL de proteínas totales del extracto se obtuvo inhibición significativa (p< 0,001) del crecimiento de S. aureus (12,64 por ciento) y E. coli (12,1 por ciento) respecto al control positivo de inhibición del crecimiento por cloranfenicol. Conclusiones: de acuerdo con los resultados de la comparación entre las soluciones, la solución salina-ácida resultó ser la más eficiente en la extracción de moléculas antibacterianas, probablemente péptidos antimicrobianos de C. muricatus.
Introduction: marine mollusks are natural reservoirs of molecules with therapeutic potential for the treatment of infectious diseases, at a time when many antibiotic-resistant strains are being described. Objective: to compare three solutions: 30 percent acetic acid, 50 percent methanol and saline-acid (NaCl 0.6 mol/L, 1 percent HCl) according to their capacities to extract molecules with antimicrobial activity from the marine mollusk Cenchritis muricatus. Methods: the three extraction solutions were used to process the biological material, and then, the obtained extracts were analyzed in terms of total protein concentration and the bacterial growth inhibition of Staphylococcus aureus and Escherichia coli strains by means of a turbidimetric bioassay using 96 well microplates in Luria-Bertani (LB) culture medium. Results: the highest total protein concentration (7.8 mg/mL) was found in the C. muricatus extract from the saline-acid solution. Additionally, 200 mg/mL of total proteins from the extract caused significant growth inhibition (p<0.001) of S. aureus (12.64 percent) and E. coli (12.1 percent) compared to the positive control of growth inhibition using chloramphenicol. Conclusions: according to these results, the saline-acid solution proved to be more efficient in extracting molecules with antibacterial activity that are likely to be antimicrobial peptides from C. muricatus.
Assuntos
Animais , Antibacterianos/isolamento & purificação , Misturas Complexas/isolamento & purificação , Gastrópodes , SoluçõesRESUMO
INTRODUCTION: Ancylostoma sp is a potentially zoonotic geohelminth. METHODS: This study aimed to evaluate in vitro the action of crude enzyme extract of Pochonia chlamydosporia (VC4) on eggs of Ancylostoma sp in 2% water-agar and in fecal cultures. RESULTS: The percentage reduction in Ancylostoma sp egg eclosion was 76.8% in Petri dishes of the treated group compared to the control group. CONCLUSIONS: The crude enzyme extract of Pochonia chlamydosporia was effective at reducing Ancylostoma sp egg eclosion and can be used as biological control of this nematode.
Assuntos
Ancylostoma/efeitos dos fármacos , Misturas Complexas/farmacologia , Hypocreales/enzimologia , Animais , Misturas Complexas/isolamento & purificação , Óvulo/efeitos dos fármacos , Controle Biológico de Vetores/métodosRESUMO
INTRODUÇÃO: Ancylostoma sp é um geo-helminto potencialmente zoonótico. MÉTODOS: O objetivo deste trabalho foi avaliar in vitro a ação do extrato bruto enzimático de Pochonia chlamydosporia (VC4) sobre ovos de Ancylostoma sp, em meio ágar-água 2 por cento e em cultura de fezes. RESULTADOS: Observou-se um percentual de redução na eclosão dos ovos de Ancylostoma sp, de 76,8 por cento na placas de Petri do grupo tratado em relação ao grupo controle. CONCLUSÕES: O extrato bruto enzimático de Pochonia chlamydosporia foi eficiente na redução da eclosão dos ovos de Ancylostoma sp, podendo ser utilizado como controlador biológico desse nematoide.
INTRODUCTION: Ancylostoma sp is a potentially zoonotic geohelminth. METHODS: This study aimed to evaluate in vitro the action of crude enzyme extract of Pochonia chlamydosporia (VC4) on eggs of Ancylostoma sp in 2 percent water-agar and in fecal cultures. RESULTS: The percentage reduction in Ancylostoma sp egg eclosion was 76.8 percent in Petri dishes of the treated group compared to the control group. CONCLUSIONS: The crude enzyme extract of Pochonia chlamydosporia was effective at reducing Ancylostoma sp egg eclosion and can be used as biological control of this nematode.
Assuntos
Animais , Ancylostoma/efeitos dos fármacos , Misturas Complexas/farmacologia , Hypocreales/enzimologia , Misturas Complexas/isolamento & purificação , Óvulo/efeitos dos fármacos , Controle Biológico de Vetores/métodosRESUMO
INTRODUCTION: marine mollusks are natural reservoirs of molecules with therapeutic potential for the treatment of infectious diseases, at a time when many antibiotic-resistant strains are being described. OBJECTIVE: to compare three solutions: 30 % acetic acid, 50 % methanol and saline-acid (NaCl 0.6 mol/L, 1 % HCl) according to their capacities to extract molecules with antimicrobial activity from the marine mollusk Cenchritis muricawus. METHODS: the three extraction solutions were used to process the biological material, and then, the obtained extracts were analyzed in terms of total protein concentration and the bacterial growth inhibition of Staphylococcus aureus and Escherichia coli strains by means of a turbidimetric bioassay using 96 well microplates in Luria-Bertani (LB) culture medium. RESULTS: the highest total protein concentration (7.8 microg/mL) was found in the C. muricatus extract from the saline-acid solution. Additionally, 200 microg/mL of total proteins from the extract caused significant growth inhibition (p< 0.001) of S. aureus (12.64 %) and E. coli (12.1 %) compared to the positive control of growth inhibition using chloramphenicol. CONCLUSIONS: according to these results, the saline-acid solution proved to be more efficient in extracting molecules with antibacterial activity that are likely to be antimicrobial peptides from C. muricatus.
Assuntos
Antibacterianos/isolamento & purificação , Misturas Complexas/isolamento & purificação , Gastrópodes , Animais , SoluçõesRESUMO
Croton cajucara is a plant found in the Amazon region and is known for its medicinal properties. The effects of the methanolic extract of the stem bark of C. cajucara (MCC) and of the isolated terpenes, trans-dehydrocrotonin (t-DCTN) and acetyl aleuritolic acid (AAA), were investigated using four isolates of Trypanosoma cruzi. In assays with trypomastigotes, the extract was more active than the isolated compounds, presenting IC(50) in the range of 10 to 50 µg/mL. The trypanocidal effect of MCC, AAA and benznidazole was significantly higher in the GLT291 and C45 strains, which were recently isolated from wild animals. MCC and AAA caused a dose-dependent inhibition of epimastigote proliferation. In assays using intracellular amastigotes, AAA and MCC reduced the percent of infection and the endocytic index after 96 h of treatment, at concentrations that were non-toxic to the host cells. MCC inhibited the trypanothione reductase pathway in both epimastigotes and trypomastigotes of all the subpopulations. The absence of AAA activity on the trypanothione reductase pathway in epimastigotes of Dm28c suggests heterogeneity of the biochemical profile between this clone and the three strains. Epimastigotes and trypomastigotes (GLT291) were treated for 24 h with MCC or AAA, and both induced alterations of the plasma membrane, while AAA-treated epimastigotes also displayed mitochondrial damage.