Development of whitening mouth rinses formulated with mushroom residues and their effect on enamel's physical properties.

Objectives: This study aimed to develop whitening mouth rinses formulated with industrial mushrooms and compare them with over-the-counter whitening mouth rinses. Materials and Methods: Formulations with black shimeji mushrooms, mushroom substrates, and mushroom stalks were developed. Bovine enamel/dentin samples were divided into 7 groups (n = 10): Colgate Luminous White, Listerine Whitening Extreme (LWE), Listerine Cool Mint (LC), mushroom extract rinse (MEC), mushroom substrate rinse (MSB), mushroom stalk rinse (MTC), and artificial saliva. Samples were stained with black tea for 6 days, and then were immersed in 100 mL of each mouth rinse twice daily for 14 days. Color parameters (CIELAB [ΔE*], CIEDE2000 [ΔE00], whiteness index for dentistry [ΔWID]) and microhardness (Knoop hardness number [KHN]) were analyzed at T1 (initial), T2 (24 hours), and T3 (7 days). Mouth rinse pH was measured, and enamel was examined using a scanning electron microscope. Data were analyzed using generalized linear models, and KHN with the generalized linear mixed model for repeated measures (p ≤ 0.05). Results: ΔE* was higher in LW and MSB groups. No significant differences were found for ΔE00 (p = 0.0982) and ΔWID (p = 0.2536). Experimental mouth rinses did not promote enamel whitening based on ΔE00 and ΔWID. LWE and LC reduced KHN and had a more acidic pH, while MEC had higher KHN at T2. MEC, MSB, and MTC had alkaline pH, not altering the tooth surface. Conclusions: Black shimeji mushrooms are promising for mouth rinse development due to their alkaline pH and non-altering effect on surface microhardness.

Postharvest of fresh white shimeji mushroom subjected to UV-C radiation.

This study aimed to evaluate the postharvest characteristics of edible fresh white shimeji mushrooms under different UV-C radiation doses. The experimental design used was fully randomized, in a 5 × 8 factorial scheme (UV-C radiation dose: 0 (control), 1, 2, 3, and 4 kJ m-2 x day of analysis), with 3 replications of 70 ± 1 g mushrooms each. After exposure to different doses, they were stored at 2 ± 0.5 °C and 60 ± 3.8 % RH. Data were subjected to permutational multivariate analysis (PERMANOVA) (p ≤ 0.05). There was no significance for interaction, nor the factor day, only for the UV-C radiation doses factor. Regarding PCA, among the doses applied, the dose of 2 kJ m-2 was effective in maintaining the quality of mushrooms with greater lightness, greater whiteness index, a greater amount of total extractable polyphenols, and total antioxidant activity. In conclusion, the dose of 2 kJ m-2 was effective in maintaining the postharvest quality of white shimeji mushrooms.

Ensayo de citotoxicidad de extractos de Pleurotusostreatus en diferentes líneas celulares para aplicaciones inmunonutricionales / Cytotoxicity assay of Pleurotus ostreatusextracts in different cell lines to immunonutritional applications / Cytotoxicity assay of Pleurotus ostreatusextracts in different cell lines to immunonutritional applications / Ensaio de citotoxicidade de extratos de Pleurotus ostreatus em diferentes linhagens celulares para aplicações imunonutricionais

Introducción: Los bioderivados propuestos como candidatos a ingredientes alimentarios suelen requerir ciertas evaluaciones para las aplicaciones inmunonutricionales Los hongos comestibles-medicinales son un surtidor de compuestos con estas potencialidades. Entre ellos, las setas Pleurotus ostreatus contienen metabolitos bioactivos, con importantes usos en la industria alimenticia y en la práctica terapéutica de la industria médico-farmacéutica. Los ensayos de citotoxicidad in vitro constituyen métodos valiosos para evaluarproductos de origen natural, como los extractos fúngicos. Objetivo: Evaluar la citotoxicidad de dos extractos obtenidos de la seta Pleurotus ostreatus en diferentes líneas celulares. Método: Se obtuvieron extractos hidrosolubles a partir del micelio y de los cuerpos fructíferos de Pleurotus ostreatus en laboratorios del Centro de Estudios de Biotecnología Industrial de la Universidad de Oriente. Se evaluó la citotoxicidad de los bioproductos por el ensayo de reducción del colorante resazurina sobre tres líneas celulares en el Laboratorio de Microbiología, Parasitología e Higiene (LMPH) de la Universidad de Amberes, Bélgica. Se utilizaron células no adherentes THP-1 (pre-monocitos de leucemia humana), células adherentes Caco-2 (epitelio de adenocarcinoma de colon humano) y células adherentes RAW 264.7 (macrófagos murinos). Resultados: Los extractos de Pleurotus ostreatus no resultaron citotóxicos para ninguna de las líneas celulares estudiadas humanas o murina, ya que no ocasionaron daños sobre la viabilidad de las célulasepiteliales del sistema gastrointestinal, nisobrelas células del sistema inmune empleadas. Conclusiones: Este resultado demuestra que ambos bioderivados fúngicos pueden ser aplicados con seguridad en estudios inmunonutricionales.(AU)

Introduction: Bioderivatives proposed as candidates for food ingredients usually require certain evaluations for immunonutritional applications. Edible-medicinal mushrooms are a source of compounds with these potentials. Among them, Pleurotus ostreatus mushrooms contain bioactive metabolites, with important uses in the food industry and in the therapeutic practice of the medical-pharmaceutical industry. In vitro cytotoxicity assays are valuable methods to evaluate products of natural origin, such as fungal extracts. Objective: To evaluate the cytotoxicity of two extracts obtained from the Pleurotus ostreatus mushroom in different cell lines. Method: Water-soluble extracts were obtained from the mycelium and fruiting bodies of Pleurotus ostreatus in laboratories of the Center for Industrial Biotechnology Studies of the Universidad de Oriente. The cytotoxicity of the bioproducts was evaluated by the resazurin dye reduction assay on three cell lines at the Laboratory of Microbiology, Parasitology and Hygiene (LMPH) of the University of Antwerp, Belgium. Non-adherent THP-1 cells (human leukemia pre-monocytes), Caco-2 adherent cells (human colon adenocarcinoma epithelium) and RAW 264.7 adherent cells (murine macrophages) were used. Results: Pleurotus ostreatus extracts were not cytotoxic for any of the human or murine cell lines studied, since they did not cause damage to the viability of the epithelial cells of the gastrointestinal system, nor to the immune system cells used. Conclusions: This result demonstrates that both fungal bioderivatives can be safely applied in immunonutritional studies.(AU)

Introdução: Bioderivados propostos como candidatos a ingredientes alimentícios geralmente requerem determinadas avaliações para aplicações imunonutricionais. Pleurotus ostreatus contêm metabólitos bioativos, com importantes utilizações na indústria alimentícia e na prática terapêutica da indústria médico-farmacêutica. Ensaios de citotoxicidade in vitro são métodos valiosos para avaliar produtos de origem natural, como extratos de fungos. Objetivo: Avaliar a citotoxicidade de dois extratos obtidos do cogumelo Pleurotus ostreatus em diferentes linhagens celulares. Método: Extratos hidrossolúveis foram obtidos do micélio e dos corpos frutíferos de Pleurotus ostreatus nos laboratórios do Centro de Estudos de Biotecnologia Industrial da Universidade de Oriente. A citotoxicidade dos bioprodutos foi avaliada pelo ensaio de redução do corante resazurina em três linhagens celulares no Laboratório de Microbiologia, Parasitologia e Higiene (LMPH) da Universidade de Antuérpia, Bélgica. Foram utilizadas células THP-1 não aderentes (pré-monócitos de leucemia humana), células aderentes Caco-2 (epitélio de adenocarcinoma do cólon humano) e células aderentes RAW 264.7 (macrófagos murinos). Resultados: Os extratos de Pleurotus ostreatus não foram citotóxicos para nenhuma das linhagens celulares humanas ou murinas estudadas, pois não causaram danos à viabilidade das células epiteliais do sistema gastrointestinal, nem às células do sistema imunológico utilizadas. Conclusões: Este resultado demonstra que ambos os bioderivados fúngicos podem ser aplicados com segurança em estudos imunonutricionais.(AU)

From purposeless residues to biocomposites: A hyphae made connection.

Biocomposites create attractive alternatives to match packing needs with available agricultural residues. Growing native fungal strains developed a mycelium biocomposite over a mixture of Peach Palm Fruit Peel Flour and Sugar Cane Bagasse Wet Dust. A methodology was proposed to analyze their main characteristics: 1) morphological, 2) chemical, and 3) biodegradability. 1) SEM analysis evidenced the structural change of the dried vs pressed material and mycelium morphology for both species. 2) The ratio lignin:carbohydrate showed that P. ostreatus degrades the cellulose-hemicellulose fraction of the substrate at a higher rate than T. elegans, and 3) the curve BMP indicated that these materials are readily biodegradable with a maximum yield of 362,50 mL biogas/g VS. An innovative tangible valorization strategy based on mass balances is also presented: from just 50 kg of peel flour, up to 1840 units can be manufactured, which could pave the way for a more sustainable future.

Possible effects of different types of agricultural wastes on food security and mushroom (Pleurotus ostreatus) production / Possíveis efeitos de diferentes tipos de resíduos agrícolas na segurança alimentar e na produção de cogumelos (Pleurotus ostreatus)

One of the new waters, and environmentally friendly agriculture initiatives in Peru is to encourage the utilization of agricultural waste, because low agricultural output is a threat to food security there. The purpose of this research was to evaluate the effect of harvest residues on the basidiocarp production of the fungus Pleurotus Ostreatus, in Acobamba-Huancavelica. The trial had a completely randomized design, and the treatments included T1, barley stubbles; T2, wheat stubbles; T3, pea stubbles; T4, broad bean stubbles; and T5, quinoa stubbles. The research was quantitative in nature, taking the form of an experiment with an applied, explanatory level of design. The recorded data was tabulated and analyzed with analysis of variance, as well as Tukey's test (α:0.05), for which the statistical software Infostat was used. The results are presented in tables and graphs for a better interpretation. As main results, it was obtained that the time (colonization), diameter (stem, pileus), length (stem) and weight (basidiocarps), present statistical differences between treatments showing significant enhancement in all parameters. Despite a numerical difference, a Tukey average comparison test revealed that there was no statistically significant difference between the averages for the variable time for fungus colonisation, suggesting that the treatment T5 in which quinoa substrate showed the greatest average. Treatment T4 in which broad bean stubbles were used gave the most low-average. In conclusion, increment in all parámeters were noted in all treatment of Pleurotus basidiocarps ostreatus under Acobamba conditions.

Uma das novas iniciativas de águas e agricultura ecológica no Peru é incentivar a utilização de resíduos agrícolas, porque a baixa produção agrícola é uma ameaça à segurança alimentar no país. O objetivo desta pesquisa foi avaliar o efeito dos resíduos da colheita na produção de basidiocarpos do fungo Pleurotus ostreatus, em Acobamba-Huancavelica. O ensaio teve um delineamento inteiramente casualizado, e os tratamentos incluíram T1, restolho de cevada; T2, soqueiras de trigo; T3, restolho de ervilha; T4, restolho de fava; e T5, restolho de quinoa. A pesquisa foi de natureza quantitativa, assumindo a forma de experimento com um nível de design aplicado e explicativo. Os dados registrados foram tabulados e analisados com análise de variância, bem como teste de Tukey (α:0,05), para o qual foi utilizado o software estatístico Infostat. Os resultados são apresentados em tabelas e gráficos para melhor interpretação. Como principais resultados, obteve-se que o tempo (colonização), diâmetro (caule, píleo), comprimento (caule) e peso (basidiocarpos) apresentam diferenças estatísticas entre os tratamentos, mostrando melhora significativa em todos os parâmetros. Apesar da diferença numérica, o teste de comparação de médias de Tukey revelou que não houve diferença estatisticamente significativa entre as médias para a variável tempo de colonização do fungo, sugerindo que o tratamento T5 em que usou o substrato quinoa, apresentou a maior média. O tratamento T4, no qual foram usadas restolhos de fava, apresentou a média mais baixa. Em conclusão, incrementos em todos os parâmetros foram observados em todos os tratamentos de Pleurotus basidiocarps ostreatus nas condições de Acobamba.

Evaluación in vivo de la pirogenicidad de bioproductos fúngicos con potencial prebiótico / In vivo pyrogenicity evaluation of fungal bioproducts with prebiotic potential / Avaliação in vivo da pirogenicidade de bioprodutos fúngicos com potencial prebiótico

RESUMEN Introducción: Los productos naturales con actividad farmacológica requieren de evaluaciones preclínicas que justifiquen su empleo sobre una base científica. El ensayo de pirógenos es una prueba dentro de la Farmacología de Seguridad que se realiza para determinar la presencia de endotoxinas y constituye un método valioso, para demostrar la seguridad de bioderivados con potencial prebiótico en el campo de la inmunonutrición. Objetivo: Evaluar la pirogenicidad de bioproductos fúngicos de Pleurotus ostreatus (extractos acuosos del micelio y cuerpos fructíferos) y un biopreparado de levadura Kluyveromyces marxianus, empleando el ensayo de pirógenos en conejos Nueva Zelanda. Método: Se ensayaron concentraciones de 1,0 y 10,0 mg/mL de cada muestra por vía endovenosa en dosis de 0,5 y 5,0 mg/kg de peso. El diseño experimental cumplió las buenas prácticas de laboratorio según lo establecido por el International Council for Laboratory Animals Science y se realizó de acuerdo a los procedimientos normalizados de trabajo del Centro de Toxicología y Biomedicina, Santiago de Cuba. Resultados: Los extractos de Pleurotus ostreatus y el biopreparado de levadura (0,5 mg/kg) no mostraron signos de pirogenicidad. En los resultados del biopreparado (5,0 mg/kg), los valores de temperatura caen en un rango de incertidumbre, según la Farmacopea de Estados Unidos (USP) y se sugirió repetir el estudio. Conclusiones: Los extractos de Pleurotus ostreatus y el biopreparado de Kluyveromyces marxianus (0,5 mg/kg) no indujeron un aumento de temperatura significativo en los animales, lo cual sugiere que en estos bioproductos no existen niveles de endotoxinas que puedan provocar pirogenicidad.

ABSTRACT Introduction: Natural products with pharmacological activity require preclinical evaluations to justify their uses scientifically. The pyrogen assay is a safety pharmacology test performed to determine the presence of endotoxins and it is a valuable method to demonstrate the bio-derivative products safety and their prebiotic potential in the field of immunonutrition. Objective: To evaluate the pyrogenicity of fungal bioproducts from Pleurotus ostreatus (aqueous extracts from mycelium and fruiting bodies) and a biopreparation from Kluyveromyces marxianus yeast, using a pyrogen assay in New Zealand rabbits. Method: Concentrations of 1.0 and 10.0 mg/mL of each sample were tested intravenously at doses of 0.5 and 5.0 mg/kg body weight. The experimental design complied with good laboratory practices as established by the International Council for Laboratory Animal Science and was carried out according to the standard work procedures of the Centro de Toxicología y Biomedicina, Santiago de Cuba. Results: Pleurotus ostreatus extracts and the yeast biopreparation (0.5 mg/kg) showed no signs of pyrogenicity. In the biopreparation results (5.0 mg/kg), temperature values fall in the uncertainty range according to the United States Pharmacopoeia (USP), and therefore it was suggested to repeat the study. Conclusions: Pleurotus ostreatus extracts and Kluyveromyces marxianus biopreparation (0.5 mg/kg) did not induce a significant temperature increase in the animals, thereby suggesting that there are no endotoxin levels in such bioproducts that could cause pyrogenicity.

RESUMO Introdução: Produtos naturais com atividade farmacológica requerem avaliações pré-clínicas que justifiquem seu uso em bases científicas. O ensaio de pirogênio é um teste dentro da Farmacologia de Segurança que é realizado para determinar a presença de endotoxinas e é um método valioso para demonstrar a segurança de bioderivados com potencial prebiótico no campo da imunonutrição. Objetivo: Avaliar a pirogenicidade de bioprodutos fúngicos de Pleurotus ostreatus (extratos aquosos do micélio e corpos de frutificação) e de uma biopreparação da levedura Kluyveromyces marxianus, utilizando o ensaio pirogênico de coelho da Nova Zelândia. Método: Concentrações de 1,0 e 10,0 mg/mL de cada amostra foram testadas por via intravenosa nas doses de 0,5 e 5,0 mg/kg de peso. O desenho experimental obedeceu às boas práticas laboratoriais estabelecidas pelo Conselho Internacional para a Ciência dos Animais de Laboratório e foi realizado de acordo com os procedimentos de trabalho padrão do Centro de Toxicologia e Biomedicina, Santiago de Cuba. Resultados: Os extratos de Pleurotus ostreatus e a biopreparação de leveduras (0,5 mg/kg) não apresentaram sinais de pirogenicidade. Nos resultados da biopreparação (5,0 mg/kg), os valores de temperatura estão dentro de uma faixa de incerteza, segundo a Farmacopeia dos Estados Unidos (USP) e foi sugerido repetir o estudo. Conclusões: Os extratos de Pleurotus ostreatus e a biopreparação de Kluyveromyces marxianus (0,5 mg/kg) não induziram um aumento significativo da temperatura nos animais, o que sugere que não há níveis de endotoxinas nesses bioprodutos que possam causar pirogenicidade.

Assessing the Biodegradation of Vulcanised Rubber Particles by Fungi Using Genetic, Molecular and Surface Analysis.

Millions of tonnes of tyre waste are discarded annually and are considered one of the most difficult solid wastes to recycle. A sustainable alternative for the treatment of vulcanised rubber is the use of microorganisms that can biotransform polymers and aromatic compounds and then assimilate and mineralise some of the degradation products. However, vulcanised rubber materials present great resistance to biodegradation due to the presence of highly hydrophobic cross-linked structures that are provided by the additives they contain and the vulcanisation process itself. In this work, the biodegradation capabilities of 10 fungal strains cultivated in PDA and EM solid medium were studied over a period of 4 weeks. The growth of the strains, the mass loss of the vulcanised rubber particles and the surface structure were analysed after the incubation period. With the white rot fungi Trametes versicolor and Pleurotus ostreatus, biodegradation percentages of 7.5 and 6.1%, respectively, were achieved. The FTIR and SEM-EDS analyses confirmed a modification of the abundance of functional groups and elements arranged on the rubber surface, such as C, O, S, Si, and Zn, due to the biological treatment employed. The availability of genomic sequences of P. ostreatus and T. versicolor in public repositories allowed the analysis of the genetic content, genomic characteristics and specific components of both fungal species, determining some similarities between both species and their relationship with rubber biodegradation. Both fungi presented a higher number of sequences for laccases and manganese peroxidases, two extracellular enzymes responsible for many of the oxidative reactions reported in the literature. This was confirmed by measuring the laccase and peroxidase activity in cultures of T. versicolor and P. ostreatus with rubber particles, reaching between 2.8 and 3.3-times higher enzyme activity than in the absence of rubber. The integrative analysis of the results, supported by genetic and bioinformatics tools, allowed a deeper analysis of the biodegradation processes of vulcanised rubber. It is expected that this type of analysis can be used to find more efficient biotechnological solutions in the future.

Effects of lignocellulolytic enzymes on the fermentation profile, chemical composition, and in situ ruminal disappearance of whole-plant corn silage.

The objective of this study was to examine the enzyme activities of an enzymatic complex produced by Pleurotus ostreatus in different pH and the effects of adding increased application rates of this enzymatic complex on the fermentation profile, chemical composition, and in situ ruminal disappearance of whole-plant corn silage (WPCS) at the onset of fermentation and 30 d after ensiling. The lignocellulolytic enzymatic complex was obtained through in vitro cultivation of P. ostreatus. In the first experiment, the activities of laccase, lignin peroxidase (LiP), manganese peroxidase, endo- and exo-glucanase, xylanase, and mannanase were determined at pH 3, 4, 5, and 6. In the second experiment, five application rates of enzymatic complex were tested in a randomized complete block design (0, 9, 18, 27, and 36 mg of lignocellulosic enzymes/kg of fresh whole-plant corn [WPC], corresponding to 0, 0.587, 1.156, 1.734, and 2.312 g of enzymatic complex/kg of fresh WPC, respectively). There were four replicates per treatment (vacuum-sealed bags) per opening time. Bags were opened 1, 2, 3, and 7 d after ensiling (onset of fermentation period) and 30 d after ensiling to evaluate the fermentation profile, chemical composition, and in situ dry matter and neutral fiber detergent disappearance of WPCS. Laccase had the greatest activity at pH 5 (P < 0.01), whereas manganese peroxidase and LiP had the greatest activity at pH 4 (P < 0.01; P < 0.01). There was no effect of the rate of application of enzymatic complex, at the onset of fermentation, on the fermentation profile (P > 0.21), and chemical composition (P > 0.36). The concentration of water-soluble carbohydrate quadratically decreased (P < 0.01) over the ensiling time at the onset of fermentation, leading to a quadratic increase of lactic acid (P = 0.02) and a linear increase of acetic acid (P = 0.02) throughout fermentation. Consequently, pH quadratically decreased (P < 0.01). Lignin concentration linearly decreased (P = 0.04) with the enzymatic complex application rates at 30 d of storage; however, other nutrients and fermentation profiles did not change (P > 0.11) with the enzymatic complex application rates. Addition of lignocellulolytic enzymatic complex from P. ostreatus cultivation to WPC at ensiling decreased WPCS lignin concentration 30 d after ensiling; however, it was not sufficient to improve in situ disappearance of fiber and dry matter.

Application of K-Means Clustering Algorithm to Commercial Parameters of Pleurotus spp. Cultivated on Representative Agricultural Wastes from Province of Guayas.

Data of the commercial parameters of Pleurotus ostreatus and Pleurotus djamor were analyzed using the data mining technique: K-means clustering algorithm. The parameters evaluated were: biological efficiency, crop yield ratio, productivity rate, nutritional composition, antioxidant and antimicrobial activities in the production of fruit bodies of 50 strains of Pleurotus ostreatus and 50 strains of Pleurotus djamor, cultivated on the most representative agricultural wastes from the province of Guayas: 80% sugarcane bagasse and 20% wheat straw (M1), and 60% wheat straw and 40% sugarcane bagasse (M2). The database of the parameters obtained in experimental procedures was grouped into three clusters, providing a visualization of the strains with a higher relation to each parameter (vector) measured.

Ligninolytic activity of the Penicillium chrysogenum and Pleurotus ostreatus fungi involved in the biotransformation of synthetic multi-walled carbon nanotubes modify its toxicity.

Multi-walled carbon nanotubes (MWCNTs) are of multidisciplinary scientific interest due to their exceptional physicochemical properties and a broad range of applications. However, they are considered potentially toxic nanoparticles when they accumulate in the environment. Given their ability to oxidize resistant polymers, mycorremediation with lignocellulolytic fungi are suggested as biological alternatives to the mineralization of MWCNTs. Hence, this study involves the ability of two fungi specie to MWCNTs biotransformation by laccase and peroxidases induction and evaluation in vivo of its toxicity using Caenorhabditis elegans worms as a model. Results showed that the fungi Penicillium chrysogenum and Pleurotus ostreatus were capable to grow on media with MWCNTs supplemented with glucose or lignin. Activities of lignin-peroxidase, manganese-peroxidase, and laccase in cultures of both fungi were induced by MWCNTs. Raman, FTIR spectroscopy, HR-TEM, and TGA analyses of the residue from the cultures of both fungi revealed structural modifications on the surface of MWCNTs and its amount diminished, correlating the MWCNTs structural modifications with the laccase-peroxidase activities in the fungal cultures. Results indicate that the degree of toxicity of MWCNTs on the C. elegans model was enhanced by the structure modification associated with the fungal ligninolytic activity. The toxic effect of MWCNTs on the in vivo model of worms reveals the increment of reactive oxygen species as a mechanism of toxicity. Findings indicate that the MWCNTs can be subject in nature to biotransformation processes such as the fungal metabolism, which contribute to modify their toxicity properties on susceptible organisms and contributing to environmental elimination.

Differential Induction Pattern Towards Classically Activated Macrophages in Response to an Immunomodulatory Extract from Pleurotus ostreatus Mycelium.

Pleurotus ostreatus mushroom preparations have been investigated because of their ability to modulate the immune function. However, there is still no consensus regarding the activation and polarizing effect on macrophages by Pleurotus-derived bioproducts. This study examined the immune-activating effect of a mycelium-derived P. ostreatus aqueous extract (HW-Pm) on macrophage functions, by means of the determination of nitric oxide (NO) production, the mRNA expression of inducible nitric oxide synthase (iNOS), Arginase-1 and FIZZ and the cytokine levels. The phagocytic activity and the activation of NF-κB in U937 reporter cells were also investigated. No cytotoxicity was observed in macrophages treated with HW-Pm (IC50 > 1024 µg/mL) by the resazurin test. HW-Pm induced high levels of NO production and iNOS expression in macrophages. In contrast, HW-Pm did not induce Arginase-1 and FIZZ mRNA expressions. The mushroom extract increased TNF-α and IL-6 production and the phagocytic function in murine macrophages. It also stimulated the activation of the NF-κB promoter. The P. ostreatus mycelium extract has a potential application as a natural immune-enhancing agent, by targeting macrophage activation towards the classically activated subset and stimulating macrophage-mediated innate immune responses.

Evaluation of functional and nutritional potential of a protein concentrate from Pleurotus ostreatus mushroom.

Edible mushrooms used as a protein-rich food may be an attractive alternative to conventional protein sources, while promoting its valorization. This work aimed to obtain a protein concentrate from a Pleurotus ostreatus mushroom flour, its characterization, and nutritional and functional properties evaluation. Methodologies applied for extraction and precipitation of protein were optimized - pH 4 and 12, respectively; and flour-solvent ratio of 1:20 w/v. The protein density was increased by 78%. P. ostreatus flour and concentrate were characterized by proximal composition. The content of total phenolic compounds in the protein concentrate decreased, leading to a positive effect on protein digestibility, while the DPPH radical scavenging activity was not significantly affected. Peptides with molecular weights from 12 to 35 kDa, with possible bioactivity, were identified by electrophoresis. Protein digestibility assessed by in vitro gastrointestinal digestion showed a 4.2-fold higher hydrolysis degree in the protein concentrate than the flour.

Evaluación in silico del efecto de compuestos fenólicos de Pleurotus ostreatus sobre la enzima 5-lipoxigenasa (5- LOX) / In silico evaluation of the effect of Pleurotus ostreatus phenolic compounds on the enzyme 5-lipoxygenase (5-LOX)

Introducción: Los hongos comestibles, en particular Pleurotus ostreatus, representan una importante fuente de metabolitos bioactivos con propiedades inmunomoduladoras, antioxidantes y antiinflamatorias. Trabajos recientes han demostrado que extractos y compuestos purificados a partir de esta seta, entre ellos, la fracción rica en fenoles, inhiben el factor nuclear kappa B(NF-κB), la cicloxigenasa (COX) y modulan cascadas de señalización relacionadas con el balance redox. De acuerdo con estos antecedentes, dichos compuestos podrían actuar, además, como inhibidores de la enzima 5- lipoxigenasa (5-LOX). Objetivo: Evaluar el efecto in silico de trece compuestos fenólicos presentes en la especie Pleurotus ostreatus sobre la enzima 5-LOX, al utilizar como compuesto de referencia la mangiferina. Métodos: El acoplamiento se llevó a cabo a través del programa AutoDock 4.2 (http://autodock.scripps.edu) y la estructura de 5 LOX se obtuvo con la base de datos de proteínas, PDB (www.wwpdb.org). Se estimaron la energía libre (ΔG), la constante de disociación (Ki) y la eficiencia de ligando (LE). Se obtuvieron los parámetros de similitud a un fármaco y los relacionados con la absorción, distribución, metabolismo, excreción y toxicidad (ADME/T) de los mejores modelos de acoplamiento. Resultados: Los mejores indicadores de ΔG y Ki, correspondieron a los ácidos homogentísico, clorogénico y gentísico, con valores de ΔG (-11,81; -12,28 y -11,67 kcal/moL) y Ki (2,19 10-9; 9,99 10-10, 2,79 10-9 M), respectivamente. La eficiencia de ligando alcanzó valores adecuados para estos tres compuestos fenólicos. El modelo de acoplamiento del ácido homogentísico mostró los mejores resultados en cuanto a la similitud a un fármaco y pruebas ADME/T. Conclusiones: El estudio in silico reveló las potencialidades de la fracción rica en fenoles de P. ostreatus, y en particular, del ácido homogentísico como inhibidor de la enzima 5 -LOX, y justifica el desarrollo de ensayos confirmativos in vitro/ in vivo que corroboren sus efectos antioxidantes y antinflamatorios(AU)

Introduction: Edible mushrooms, Pleurotus ostreatus in particular, are an important source of bioactive metabolites with immunomodulatory, antioxidant and anti-inflammatory properties. Recent studies have shown that extracts and compounds purified from this mushroom, among them the phenol-rich fraction, inhibit nuclear factor kappa B (NF-κB), cyclooxygenase (COX), and modulate signaling cascades related to redox balance. According to these antecedents, such compounds could also act as inhibitors of the enzyme 5-lipoxygenase (5-LOX). Objective: Evaluate the in silico effect of 13 phenolic compounds present in the species Pleurotus ostreatus on the enzyme 5-LOX using mangiferin as reference compound. Methods: Docking was carried out with the software AutoDock 4.2 (http://autodock.scripps.edu) and the 5-LOX structure was obtained with the protein database PDB (www.wwpdb.org). Estimation was performed of free energy (ΔG), dissociation constant (Kd) and ligand efficiency (LE). Drug-likeness parameters were obtained, as well as those related to absorption, distribution, metabolism, excretion and toxicity (ADMET) of the best docking models. Results: The best ΔG and Kd indicators were homogentisic, chlorogenic and gentisic acids, with ΔG and Kd values of -11.81, -12.28, -11.67 kcal/mol, and 2.19 10-9, 9.99 10-10, 2.79 10-9 M, respectively. Ligand efficiency achieved adequate values for these three phenolic compounds. The docking model for homogentisic acid showed the best results in terms of drug likeness and ADMET tests. Conclusions: The in silico study revealed the potential of the phenol-rich fraction of P. ostreatus, homogentisic acid in particular, as an enzyme 5-LOX inhibitor, and justifies the development of confirmatory in vitro / in vivo assays to corroborate its antioxidant and anti-inflammatory effects(AU)

Secondary Metabolites and Antioxidant Activity of the Solid-State Fermentation in Apple (Pirus malus L.) and Agave Mezcalero (Agave angustifolia H.) Bagasse.

Solid-state fermentation (SSF) is used in enzyme and antibiotic production, bioethanol and biodiesel as an alternative energy source, biosurfactants with environmental goals, and the production of organic acids and bioactive compounds. The present project determined the quantity of secondary metabolites and the antioxidant activity of the extracts obtained by the solid-state fermentation of apple and agave mezcalero bagasse over 28 days, inoculated with the Pleurotus ostreatus strain. The extraction was carried out with three solvents: acetone and water (80:20 v/v), 100% methanol and 100% water. The results showed a higher presence of phenolic compounds, flavonoids, total triterpenes and antioxidant activity in the apple bagasse from the SSF on day 21 in the extract of acetone and water (80:20 v/v), 100% methanol and aqueous; while the agave bagasse showed a significant presence of phenolic compounds and flavonoids only in the aqueous extract. In conclusion, the presence of secondary metabolites exhibiting antioxidant activities from the solid-state fermentation in the residues of the cider and mezcal industry is an alternative use for wasted raw material, plus, it reduces the pollution generated from the agroindustrial residues.

Reduction of Salt and Fat in Frankfurter Sausages by Addition of Agaricus bisporus and Pleurotus ostreatus Flour.

The reduction of fat and salt and the incorporation of fiber-rich compounds in frankfurters is a trend to improve their nutritional profile. The objective of this study was to evaluate the partial replacement of 30 and 50% of pork backfat and 50% of salt by adding edible mushroom flour (2.5 and 5%) from Agaricus bisporus (Ab) and Pleurotus ostreatus (Po) on physicochemical, microbiological and sensory properties of frankfurters sausages during cold storage. The addition of flours increased the moisture, and the dietary fiber contents in frankfurters, keeping the amino acid profile. Lipid oxidation remained under acceptable values despite not antioxidant effect was observed by mushrooms flours. Only spore-forming bacteria were found during cold storage. Color and texture was modified by addition of mushroom, being the Ab samples darker, while Po flour addition resulted in softer and less cohesive sausages. Although lower color, flavor, and taste scores were given to the mushroom samples than the control, they ranked in the acceptable level confirming that the inclusion of 2.5 and 5% of Ab and Po flours in fat- and salt-reduced frankfurter sausages resulted a feasible strategy to enhance the nutritional profile these products.

Micetismos: Parte 4: Síndromes tempranos con síntomas complejos / Mushroom poisonings. Part 4: early-onset syndromes with complex symptoms / Intoxicações por cogumelos. Parte 4: Síndromes precoces com sintomas complexos

En esta Parte 4 de la serie de cuatro artículos sobre micetismos se analizan los síndromes que se caracterizan por presentar un período de latencia muy corto, con la aparición de síntomas complejos en menos de 6 horas después de la ingestión de los macromicetos. Se discuten los siguientes micetismos: 1) Toxíndrome muscarínico o colinérgico periférico por especies de Inocybe y Clitocybe. 2) Toxíndrome inmunohemolítico o hemolítico por Paxillus. 3) Toxíndrome neumónico alérgico por Lycoperdon perlatum y por Pholiota nameko. 4) Toxíndrome panterínico o neurotóxico glutaminérgico por compuestos isoxazólicos o síndrome pantherina/muscaria. 5) Toxíndrome coprínico o cardiovascular. 6) Toxíndrome neurotóxico alucinogénico por psilocibina y derivados indólicos. 7) Toxíndrome psicotrópico por estirilpironas y gimnopilinas de Gymnopilus spectabilis o G. junonius. 8) Toxíndrome agudo de rabdomiólisis por Russula subnigricans. 9) Toxíndrome cianogénico por Marasmius oreades. 10) Toxíndrome inmunosupresor por tricotecenos macrocíclicos de Podostroma cornu-damae. 11) Toxíndrome hemolítico debido a ostreolisina de Pleurotus ostreatus y especies relacionadas. Se analizan los síntomas, las toxinas involucradas, los mecanismos de acción, cuando se conocen, y las especies causantes de los micetismos.

This Part 4 of the series of four articles on mushroom poisonings refers to early-onset syndromes, which are characterized by a very short latency period, and the appearance of complex symptoms in less than 6 hours after mushroom ingestion. The following mycetisms are discussed, (1) Peripheral cholinergic, or muscarinic syndrome due to Inocybe and Clitocybe species. (2) Immunohaemolytic or haemolytic syndrome by Paxillus. (3) Allergic pneumonic syndrome due to Lycoperdon perlatum, and Pholiota nameko. (4) Glutaminergic neurotoxic, or pantherinic syndrome by isoxazole compounds or pantherina/muscaria syndrome. (5) Coprinic or cardiovascular syndrome. (6) Hallucinogenic neurotoxic syndrome due to psilocybin and indole derivatives. (7) Psychotropic syndrome by styrylpirones and gymnopilins of Gymnopilus spectabilis or G. junonius. (8) Rhabdomyolysis acute syndrome due to Russula subnigricans. (9) Cyanogenic syndrome by Marasmius oreades. (10) Immunosuppressive syndrome by macrocyclic trichothecenes of Podostroma cornu-damae. (11) Haemolytic syndrome due to ostreolisine of Pleurotus ostreatus, and related species. The symptoms, toxins involved, mechanisms of action, when known, and the species of mushrooms responsible for the mycetisms are analyzed.

Nesta parte 4 da série de quatro artigos sobre intoxicação por cogumelos são analisadas síndromes que se caracterizam por apresentar um período de latência muito breve, com aparecimento de sintomas complexos em menos de 6 horas após a ingestão dos macromicetos. As seguintes intoxicações com cogumelos são discutidas: (1) Toxíndrome muscarínico ou colinérgico periférico por espécies de Inocybe e Clitocybe. (2) Toxíndrome imuno-hemolítica ou hemolítica por Paxillus. (3) Toxíndrome pneumônica alérgica por Lycoperdon perlatum e por Pholiota nameko. (4) Toxíndrome panterínica ou neurotóxica glutaminérgica por compostos isoxazólicos ou síndrome pantherina/muscaria. (5) Toxíndrome coprínica ou cardiovascular (6) Toxíndrome neurotóxico-alucinogênica por psilocibina e derivados indólicos. (7) Toxíndrome psicotrópica por estirilpironas e gimnopilinas de Gymnopilus spectabilis ou G. junonius. (8) Toxíndrome aguda de rabdomiólise por Russula subnigricans. (9) Toxíndrome cianogênica por Marasmius oreades. (10) Toxíndrome imunossupressora por tricotecenos macrocíclicos de Podostroma cornu-damae. (11) Síndrome hemolítica por ostreolisina de Pleurotus ostreatus e espécies relacionadas. São analisados os sintomas, as toxinas envolvidas, os mecanismos de ação, quando conhecidos, e as espécies de cogumelos responsáveis pelas intoxicações.

Use of peanut waste for oyster mushroom substrate supplementation-oyster mushroom and peanut waste.

The aim of the research was to verify the influence of macro and micronutrients present in the peanut waste (hulls and nuts) for supplementation of Pleurotus ostreatus substrate. The raw materials for base substrate preparation were Brachiaria dictyoneura, sugarcane bagasse (bulk material), rice and wheat bran, calcitic limestone, and gypsum. The following supplement formulations were used as treatments: (1) 100% peanut hulls, (2) 80% peanut hulls + 20% nuts, (3) 60% peanut hulls + 40% nuts, (4) 40% peanut hulls + 60% nuts, (5) 20% peanut hulls + 80% nuts, and (6) 100% nuts. A commercial supplement was also used as an additional treatment. The supplementation was done at spawning using the rates of 1% and 2% wet weight of the substrate. Positive correlations amongst yield and N content, and weight of mushroom and P and K content were verified with 1% supplement. A positive correlation between yield and Cu content, and a negative correlation between yield and Mn content were observed with 2% supplement. The use of peanut waste can be used as supplement for the production of P. ostreatus increasing biological efficiency up to 61%. A better combination can be reached with 20% peanut hulls + 80% nuts or 100% nuts. The addition of 2% supplement in the substrate provided greater yield than 1%.

Combined fungal and photo-oxidative Fenton processes for the treatment of wood-laminate industrial waste effluent.

The present study reports on the remediation of an effluent from the wood-laminate industry using Pleurotus ostreatus EB 016 in combination with photo-Fenton oxidation. Fermentation of the effluent with P. ostreatus EB-016 was carried out in agitated flasks to evaluate the influence of pH, and concentrations of carbon and nitrogen sources by a multivariate approach. Subsequently, bioassays was conducted in an air-lift bioreactor using the pre-optimized conditions. In addition, photo-Fenton oxidative treatment was employed to degrade recalcitrant compounds, and the ecotoxicity of the effluents were evaluated using Escherichia coli as a biological model. The crude effluent presented high contents of total phenolics (1,220 mg/L), solids (18.45 g/L) and color intensity (8,333 CU), besides high values of chemical (COD 2,477 mg O2./L) and biochemical (BOD5, 8,450 mg O2/L) oxygen demand. Another feature was the high inhibition on Escherichia coli (71%). Reduction of 64% COD was obtained under optimized conditions (pH5.7, 7.5 g/L sucrose, 4.0 g/L ammonium nitrate) in agitated flasks after 10 days treatment. In the air-lift reactor, 50.6% COD and 29.9% total phenols were removed after 10 days. Combination of biotreatment with photo-Fenton oxidation resulted in removal of 99.2% COD and 92.2% phenolics and absence of inhibition on Escherichia coli.

Stability of immobilized laccase on Luffa Cylindrica fibers and assessment of synthetic hormone degradation.

In this work were studied the pH, thermal, and storage stability of free and immobilized laccases. Enzymes were produced by Pleurotus ostreatus on potato dextrose (PD) broth and potato dextrose modified (PDM) broth, and immobilized using Luffa cylindrica fibers as support. Both free and immobilized enzymes were assessed on their respective enzymatic activities and for 17α-ethinylestradiol (EE2) degradation. The optimum pH conditions concerning laccase activity ranged from 3.6 to 4.6, while temperature ranged between 30 °C and 50 °C for both free and immobilized enzyme. Laccase produced using PD broth presented greater storage stability and thermal stability than that of PDM. Best EE2 removals were of 79.22% and 75.00% for the free and immobilized enzymes, respectively. Removal rates were assessed during 8 h at pH 5. The removal of 17α-ethinylestradiol was stabilized in the fourth cycle of use. Results imply that immobilization promoted stability towards pH and temperature variations, although media played a decisive role in the enzymatic activity. Both free and immobilized laccases of P. ostreatus were able to degrade EE2, whereas immobilized laccase in PDM medium presented possible reuse applicability, albeit removal was not optimal when compared to other reports.

Exploring oyster mushroom (Pleurotus ostreatus) substrate preparation by varying phase I composting time: changes in bacterial communities and physicochemical composition of biomass impacting mushroom yields.

AIMS: To investigate characterization of the bacterial community composition and functionality and their impact on substrate biodegradation as well as mushroom yield. METHODS AND RESULTS: Bacterial diversity, composition and functionality were accessed by DNA-derived analysis for a sugarcane straw-based substrate composted for either 5, 10 or 15 days. In addition, carbon and nitrogen losses, carbohydrate conversion and mushroom yields were measured for the different treatments. Changes were observed in the bacterial community diversity and composition after the process started, but not during the composting process itself. Following phase I, Acinetobacter sequences were recovered in high numbers, and selected genes associated with nitrogen metabolism and lignocellulose deconstruction were mapped. Substrate physicochemical composition showed elevated carbon and nitrogen losses after 10 and 15 days of phase I with reductions in mushroom yield. CONCLUSIONS: Acinetobacter species appear to play an important role in substrate degradation processes, and a 5-day phase I period showed a significant higher mushroom yield compared to composting for either 10 or 15 days. SIGNIFICANCE AND IMPACT OF THE STUDY: This study confers a better understanding of the bacterial community manipulation during the substrate preparation and their influence in substrate selectivity for the Pleurotus ostreatus cultivation.