Estudo comparativo de diferentes cogumelos comestíveis comercializados no Brasil: ação contra radicais livres, espécies reativas e atividade antibacteriana

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Taxonomia de ácaros Rhodacaridae (Acari: Mesostigmata) e controle biológico de moscas Sciaridae (Diptera : Sciaridae) com ácaros predadores Mesostigmata em cultivo de cogumelos

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Phenol oxidation by mushroom waste extracts: a kinetic and thermodynamic study

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Yield of different white button strains in sugar cane by product-based composts

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Cadmium determination in Lentinus edodes mushroom species

Many studies have drawn attention to the occurrence and concentration of toxic elements found in the fruiting body of mushrooms. Some edible mushroom species are known to accumulate high levels of inorganic contaminants, mainly cadmium, mercury, and lead. There are about 2,000 known edible mushroom species, but only 25 of them are cultivated and used as food. In Brazil, the most marketed and consumed mushroom species are Agaricus bisporus, known as Paris champignon, Lentinus edodes, or Shitake and Pleurotus sp, also called Shimeji or Hiratake. In this study, the concentration of cadmium was determined in Lentinus edodes mushrooms from different cities in São Paulo state and some samples imported from Japan and China. The analyses were performed by graphite furnace atomic absorption spectrometry after HNO3-H2O2 digestion. The results showed a lower concentration of Cd in the mushrooms cultivated in São Paulo (0.0079 to 0.023 mg.kg-1 in natura) than that of the mushrooms cultivated abroad (0.125 to 0.212 mg.kg-1 in natura). Although there is no tolerance limit for Cd in mushrooms in Brazil, the results show that Lentinus edodes mushrooms can be safely consumed.

Nuove tecniche per l'allevamento massale di Ditteri del genere Coenosia Meigen

The species of the genus Coenosia Meigen are polyphagous predators both in the larval and in the adult stage. In Europe five Coenosia species of the ‘tigrina group’ are naturally present in greenhouses, where they can establish for long periods. As their wide preys range includes important pests of protected crops, as Aleyrodidae, Sciaridae and Agromyzidae, Coenosia species are considered promising potential biological control agents. A method for rearing Coenosia species in vivo was developed for the first time in 1993 in Germany, where C. attenuata, C. strigipes and C. humilis were bred on Bradysia paupera (Diptera Sciaridae), reared on Fusarium spp. cultivated on wood fibre. Although this method was partially simplified afterwards, it is still too complex and expensive for a mass production. This research aimed at simplifying this rearing procedure and making it cheaper, in the perspective of an eventual mass production. Studies on potential preys were conducted, to determine their suitability for C. attenuata larvae and adults and to develop rearing methods. Biology and rearing methods of Bradysia paupera Tuomikoski, Scatella stagnalis Fallén and Drosophila melanogaster Meigen (Diptera: Sciaridae, Ephydridae, Drosophilidae) were compared. B. paupera resulted the most suitable prey for rearing C. attenuata in vivo. The Sciarid fly was effectively reared on damp coconut fibre with fresh Agaricus bisporus (J.E. Lange) Pilát, thus simplifying the existing method. After preliminary trials with different potential preys, attempts to rear C. attenuata in vivo on B. paupera and D. melanogaster were made. The best results were obtained with B. paupera, reared on coconut fibre and A. bisporus, but the method needs further improvement. Trials of in vitro rearing of C. attenuata were also made: as no specific diet for Coenosia species is reported in literature, different potentially suitable media were tested. Among these, a specific diet for Diptera Tachinidae resulted a good starting point for further studies and improvements. The biology of C. attenuata adults captured in greenhouses was also studied, by observing both groups and isolated individuals. Data on lifespan, daily number of preys per adult, daily number of laid eggs and hatching rate were recorded, and the effects of different foods on these parameters were analyzed. The following foods were compared: D. melanogaster adults only, as preys for C. attenuata; D. melanogaster adults and a water-honey solution; the water-honey solution only. Honey resulted an effective food integration for C. attenuata, increasing lifespan and the number of egg laying females. It is possible that in greenhouses Coenosia adults complete the preys diet with nectar and/or honeydew. Moreover, the integration with honey reduced the daily preys consumption. This may allow to prevent cannibalism among Coenosia adults in the rearing conditions, where high population densities are required. A survey of the Coenosia species naturally present in Lombardy greenhouses was conducted. The species C. attenuata, C. strigipes, C. tigrina and C. atra were detected. C. attenuata resulted the most common, recorded in most greenhouses and for consecutive years. Besides, the presence of potential preys, weeds and the crops were recorded in each greenhouse. Nevertheless, it is difficult to determine the relation between these parameters and the presence of Coenosia species.

Produzione di enzimi da scarti agro-alimentari tramite Pleurotus ostreatus

Nel corso del tirocinio di tesi si sono studiate nuove metodologie per la produzione di enzimi idrolitici per matrici lignocellulosiche vegetali di scarto. In primis è stato valutato un nuovo metodo di produzione enzimatica utilizzando il fungo basidiomicete Pleurotus ostreatus all’interno di un fermentatore in stato solido (SSF) movimentando periodicamente il substrato mediante un'estrusione meccanica e confrontando i risultati con esperimenti analoghi ma privi di estrusione. In seguito si è valutata l’attività enzimatica prodotta dal fungo Agaricus bisporus (il comune Champignons) cresciuto tramite una fermentazione in stato solido priva di qualsiasi movimentazione. Infine gli estratti enzimatici ricavati dalle prove precedenti sono stati utilizzati allo scopo di idrolizzare matrici vegetali di scarto provenienti dall’industria cerealicola e viti-vinicola. I risultati del lavoro risultano promettenti e si osserva come sia gli estratti ricavati da fermentazioni su stato solido dinamiche (con Pleurotus) che quelle su stato solido statiche (con Agaricus) sono in grado di favorire l’idrolisi e la degradazione delle matrici vegetali favorendo la fuoriuscita di componenti di interesse come zuccheri riducenti e polifenoli.

Über die Funktion und Struktur der Tyrosinase aus Streptomyces antibioticus

Für die Aufklärung der chemisch anspruchsvollen Monophenolase-Reaktion von Tyrosinasen wurde ein System entwickelt, um das Zielprotein aus dem Bakterium Streptomyces antibioticus in großen Mengen und mit hoher Reinheit zu isolieren. Zudem konnte ein hypothetischer Reaktionsmechanismus für die Monophenolase- und die Diphenolase-Aktivität der Tyrosinase formuliert werden. Die beiden Reaktionen der S. antibioticus-Tyrosinase wurden kinetisch analysiert und auf diesem Weg die Aktivität des Enzyms mit jener der sehr gut charakterisierten Tyrosinase aus dem Pilz Agaricus bisporus verglichen. Hierbei wurden signifikante Unterschiede festgestellt, die auf die verschiedenartigen Proteinstrukturen zurückgeführt wurden. Auch konnte gezeigt werden, dass einige sekundäre Pflanzenstoffe, die vor allem in Wein zu finden sind und in ihrer chemischen Struktur den Tyrosinasesubstraten ähnlich sind, die Aktivität dieses Enzyms maßgeblich beeinflussen. Das O2-Transportprotein Hämocyanin aus der Vogelspinne Eurypelma californicum, das wie die Tyrosinase zu der Familie der Typ-3-Kupferproteine gehört, ist nach chemischer Aktivierung zur Phenoloxidase zur enzymatischen Quervernetzung von Proteinen fähig. Die Tatsache, dass diese Quervernetzung auch das Hämocyanin selbst betrifft, sowie der erfolgreiche Nachweis von Hämocyanin in der Kutikula des genannten Organismus, legen die Vermutung nahe, dass die physiologische Funktion von Hämocyanin im Rahmen der Sklerotisierung des Exoskeletts in einer aktiven und passiven Beteiligung an Gerbungsprozessen im Integument besteht.

Poder antioxidante de cogumelos comestíveis

Os cogumelos comestíveis são apreciados em todo o mundo não só pela sua textura e sabor, também pelas suas propriedades nutricionais e funcionais. Estas propriedades devem-se ao facto dos cogumelos possuírem na sua constituição compostos bioativos, nomeadamente, compostos fenólicos, tocoferóis, ácido ascórbico e carotenoides, que pelos seus efeitos no organismo estão associados à prevenção de doenças e promoção da saúde. O presente trabalho pretendeu comparar três espécies de cogumelos silvestres (Boletus edulis, Lactarius deliciosus e Macrleptiota procera) com cogumelos de cultura (Agaricus bisporus e Pleurotus ostreatus) relativamente à sua composição em macronutrientes, à sua capacidade antioxidante e ao seu teor em fenóis totais. A escolha destas espécies fundamentou-se no seu elevado consumo pela população.As amostras foram analisadas, quanto à composição de macronutrientes (humidade, proteínas, lípidos, cinzas e hidratos de carbono). Constatou-se que os cogumelos, independentemente da sua origem (selvagens ou de cultura), são maioritariamente constituídos por água com valores que oscilam entre os 76,6% -93,4% e hidratos de carbono (82 g% - 92 g% de matéria seca). Possuem ainda, quantidades baixas de gordura (0,8 g% - 1,5 g% de matéria seca). Já os valores de proteína, obtidos oscilaram entre 1,4 g% - 4,9 g% de matéria seca, apresentando os cogumelos silvestres valores mais elevados deste macronutriente. O teor de minerais totais foi variável dependendo do local onde as amostras se colheram. A avaliação da composição fenólica e da capacidade antioxidante foi feita em extratos de metanol: ácido acético (98:2) e de acetona: água (60:40). Para determinação dos compostos fenólicos recorreu-se ao método de Folin Cioacalteau, tendo-se utilizado os métodos do DPPH (radical 2,2-defenil -1- picrilhidrazilo) e do ABTS [ácido 2,2`-azino-bis (3-etilbenzotiazolina-6-sulfónico)] para análise da atividade antioxidante.Para todas as espécies analisadas o conteúdo em compostos fenólicos totais foi mais elevado para o extrato metanólico. Os cogumelos de cultura apresentavam teores de compostos fenólicos mais elevados do que os silvestres, mostrando o Pleurotus ostreatus os valores mais elevados (1,5 mg EAG/g).Em ambas as metodologias utilizadas para avaliação da capacidade antioxidante o extrato metanólico apresentou valores mais elevados tendo as espécies silvestres exibido valores superiores aos de cultura, com particular destaque para o Boletus edulis e para o Macroleptiota procera. Em conclusão, os resultados obtidos demonstram que os cogumelos são uma boa fonte de proteína e de compostos com capacidade antioxidante, apresentando variações entre espécies com destaque para os cogumelos silvestres.

Molecular dynamics simulations of the hydrophobin SC3 at a hydrophobic/hydrophilic interface

Hydrophobins are small (similar to 100 aa) proteins that have an important role in the growth and development of mycelial fungi. They are surface active and, after secretion by the fungi, self-assemble into amphipathic membranes at hydrophobic/hydrophilic interfaces, reversing the hydrophobicity of the surface. In this study, molecular dynamics simulation techniques have been used to model the process by which a specific class I hydrophobin, SC3, binds to a range of hydrophobic/ hydrophilic interfaces. The structure of SC3 used in this investigation was modeled based on the crystal structure of the class II hydrophobin HFBII using the assumption that the disulfide pairings of the eight conserved cysteine residues are maintained. The proposed model for SC3 in aqueous solution is compact and globular containing primarily P-strand and coil structures. The behavior of this model of SC3 was investigated at an air/water, an oil/water, and a hydrophobic solid/water interface. It was found that SC3 preferentially binds to the interfaces via the loop region between the third and fourth cysteine residues and that binding is associated with an increase in a-helix formation in qualitative agreement with experiment. Based on a combination of the available experiment data and the current simulation studies, we propose a possible model for SC3 self-assembly on a hydrophobic solid/water interface.

Ethanolic extracts from three commercial edibe mushrooms: determination of ergosterol contente and evaluation of bioactives properties

Mushrooms are rich in several bioactive metabolites among them are phenolic compounds, terpenoids, polysaccharides, lectins, and steroids including mycosterols, namely ergosterol [1]. Ethanolic extracts prepared by maceration of several mushroom species have been recently described as having antiinflammatory properties [2]. In the present work, ethanolic extracts of Agaricus bisporus L., Lentinus edodes (Berk.) Pegler and Pleurotus ostreatus (Jacq. ex Fr.) P.Kumm., purchased from a local supermarket in the Northeast of Portugal, were obtained by Soxhlet and chemically characterized in terms of ergosterol content by HPLC-UV. The antioxidant properties of these extracts were evaluated through DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity (RSA), reducing power (RP), p. carotene bleaching inhibition (CBI) and lipid peroxidation inhibition in TBARS (thiobarbituric acid reactive substances) assay (LPI); the antioxidant activity of ergosterol was also evaluated by the DPPH assay. The anti-inflammatory activity of the same extracts and ergosterol was evaluated in LPS (lipopolysaccharide) stimulated RAW 264.7 macrophages, through the inhibition of NO production. A. bisporus revealed the highest content in ergosterol (44.8 ± 0.4 mg/ g extract) followed by P. ostreatus (34 ± 3 mg/ g extract) and finally L. edodes (8.9 ± 0.1 mg/ g extract). A. bisporus showed the highest RSA, RP and CBI (EC50 values= 7.0 ± 0.8, 2.3 ± 0.1 and 1.4 ± 0.1 mg/mL, respectively), while L. edodes presented the highest LPI (2.5 ± 0.1 mg/mL ); ergosterol revealed higher RSA (0.46±0. 0 I mg/mL) than the extracts. Concerning the anti-inflammatory potential, the most efficient species was L. edodes (lC50 value = 164 ± 16 J.lg/mL), followed by A. bisporus (185 ± 16 J.lg/mL) and finally P. ostreatus (290 ± 10 J.lg/mL). However, ergosterol presented lower activity (338 ± 23 J.lg/mL) due to its low solubility in the culture medium. The higher antioxidant properties displayed by A. bisporus can be related with its higher ergosterol content, while in the anti-inflammatory activity this relation cannot be established also due to the low solubility of ergosterol in the cells culture medium, decreasing the ergosterol availability. More studies are being conducted regarding the ergosterol solubility. Several compounds have been implicated in the bioactivity of mushrooms and in this study we have found that ergosterol can give an important contribution.

Interaction of polysaccharide-protein complex from Agaricus blazei with Langmuir and Langmuir-Blodgett films of phospholipids

The use of natural substances in health applications may be hampered by the difficulties in establishing the mechanisms of action, especially at molecular-level. The protein-polysaccharide complex extracted from the mushroom Agaricus blazei Murill, referred to as CAb, has been considered for treating various diseases with probable interaction with cell membranes. In this study, we investigate the interaction between CAb and a cell membrane model represented by a Langmuir monolayer of dimyristoyl phosphatidic acid (DMPA). CAb affects the structural properties of DMPA monolayers causing expansion and increasing compressibility. In addition, interaction with DMPA polar heads led to neutralization of the electrical double layer, yielding a zero surface potential at large areas per molecule. CAb remained at the interface even at high surface pressures, which allowed transfer of Langmuir-Blodgett (LB) films onto solid supports with the CAb-DMPA mixture. The mass transferred, according to quartz crystal microbalance (QCM) measurements, increased linearly with the number of deposited layers. With UV-vis absorption, fluorescence and FTIR spectroscopies, we confirmed that the LB films contain polysaccharides, proteins and DMPA. Therefore, the CAb biological action must be attributed not only to polysaccharides but also to proteins in the complex. (C) 2008 Elsevier Inc. All rights reserved.

Mechanism of anticlastogenicity of Agaricus blazei Murill mushroom organic extracts in wild type CHO (K-1) and repair deficient (xrs5) cells by chromosome aberration and sister chromatid exchange assays

Agaricus blazei Murill is a medicinal mushroom native to Brazil. The present work assessed the clastogenic and anticlastogenic potential of organic extracts (ethanol and chloroform/methanol) from the lineage AB97/11 in chinese hamster CHO-K-1 (wild type) and CHO-xrs5 (repair deficient) cells using the chromosome aberration (CA) and sister chromatid exchange (SCE) assays. In these experimental conditions were observed: (a) anticlastogenic effect at concentrations of 0.06 and 0.09% of the EtOH extract and at the 0.03 and 0.06% concentrations of the C/MetOH extract in CHO-K-1; (b) absence of protector effect on CHO-xrs5 cells; and (c) absence of protector effect in the SCE assay. These results indicate that organic extracts of A. blazei lineage AB97/11 present bio-antimutagenic type protective activity. (C) 2003 Elsevier B.V. B.V. All rights reserved.

Production of Agaricus blazei ss. Heinemann (A. brasiliensis) on different casing layers and environments

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Genetic characterization of isolates of the basidiomycete Agaricus blazei by RAPD

Determinou-se a divergência genética entre cinco isolados do fungo Agaricus blazei pela técnica de RAPD. Dos cinco isolados três não apresentaram qualquer divergência, sendo caracterizados como isolados de uma mesma origem, embora obtidos em locais diferentes do país. Outros dois isolados mostraram-se diferentes do primeiro grupo e divergentes entre si.