Tannic extract potential as natural wood preservative of Acacia mearnsii

ABSTRACT High toxicity of the preservatives most frequently used in wood treatment and the resulting risks of handling pose a threat to small producers and to the environment. In an attempt to mitigate these problems, the present study was conducted with the objective of evaluating the preservative effect of tannic extract on biodeterioration of Acacia mearnsii wood. For this purpose, untreated and preserved specimens, some with tannin extract and some with a preservative mixture based on CCB (Chromated Copper Borate), were submitted to accelerated rotting trials with the fungus that causes white rot (Pycnoporus sanguineus) for 16 weeks. The evaluations were made with a basis on weight loss and chemical components analysis, and they showed that the natural resistance of Acacia wood is moderate when exposed to the white rot fungus. The tannin concentrations showed similar effects to those of the CBB mixture in all evaluations, i.e., they significantly increased the biological resistance of the material, which started to be classified as very resistant to the fungus. Overall, the results suggest that tannin can be considered as a potential natural preservative product.

Produção de lacase de Pycnoporus sanguineus em meio de cultivo à base de melaço soja / Laccase production by Pycnoporus sanguineus in culture media with soybean molasses / Producción de lacasa de Pycnoporus sanguineus en medio de cultivo con melaza de soja

Lacases são polifenol oxidases que utilizam a capacidade redox de íons cobre para reduzir oxigênio a água e oxidar um substrato fenólico. A síntese e secreção de lacases de basidiomicetos dependem de vários fatores como os nutrientes presentes no meio de cultura. Visando à produção de lacase, Pycnoporus sanguineus foi cultivado em meio contendo melaço de soja como única fonte de carbono, ureia como fonte de nitrogênio suplementar em diferentes concentrações (0,6; 1,2; 2,4; 4,8 e 9,6 g/L de nitrogênio) e diferentes concentrações de CuSO4 (0, 150, 200, 250 e 300 µM). O extrato enzimático produzido nas melhores condições de cultivo foi utilizado para a descoloração dos corantes remazol azul brilhante R (antraquinona), amarelo 145, preto 5, vermelho 195 (azo) e verde malaquita (trifenilmetano). As concentrações de nitrogênio não afetaram a produção de lacase, exceto a maior concentração (9,6 g/L) que reduziu a atividade enzimática. A adição de cobre ao meio de cultivo (150 µM) aumentou a atividade de lacase em 112%. A maior atividade de lacase (~34300 U/L) promoveu a descoloração dos corantes remazol azul brilhante R (67,5%) e verde malaquita (28,3%) em 24h, sendo os corantes azo descoloridos apenas parcialmente. Concluiu-se que o melaço de soja é um resíduo agroindustrial adequado para produção de lacase de P. sanguineus com potencial para degradação de corantes.

Laccases are multicopper oxidases using the redox ability from copper ions to reduce oxygen to water, while oxidizing a phenolic substrate. Laccase synthesis and secretion in basidiomycetes depend on the conditions provided and on the nutrients present in the culture medium. Pycnoporus sanguineus was cultivated in medium containing soybean molasses as the sole carbon source, with urea as the source of supplemental nitrogen at different concentrations (0.6, 1.2, 2.4, 4.8 and 9.6 g/L nitrogen), and different CuSO4 concentrations (0, 150, 200, 250 and 300 µM). The enzymatic extract produced under the best cultivation conditions was used for the depigmentation of remazole brilliant blue R (anthraquinone), yellow 145, black 5, red 195 (azo) and malachite green (triphenylmethane). Nitrogen concentrations did not affect laccase production, except for the higher concentration (9.6 g/L) reducing enzymatic activity. The addition of copper to the culture medium (150 µM) increased laccase activity by 112%. The highest laccase activity (~34300 U/L) promoted the depigmentation of remazol brilliant blue R (67.5%) and malachite green (28.3%) dyes in 24 hours. Azo dyes were only partially discolored. Therefore, it can be considered that soybean molasses is an agro-industrial byproduct suitable for the production of P. sanguineus laccase with potential for dye degradation.

Lacasas son polifenoles oxidasas que utilizan la capacidad redox de iones de cobre para reducir el oxígeno del agua y oxidar un sustrato fenólico. La síntesis y secreción de lacasas de basidiomicetos dependen de las condiciones como los nutrientes presentes en el medio de cultura. Buscando la producción de lacasa, se cultivó Pycnoporus sanguineus en medio que contenía melaza de soja como única fuente de carbono, urea como fuente de nitrógeno suplementar a diferentes concentraciones (0,6, 1,2, 2,4, 4,8 y 9,6 g/L de nitrógeno) y diferentes concentraciones de CuSO4 (0, 150, 200, 250 y 300 µM). El extrato enzimático producido en mejores condiciones de cultivo ha sido utilizado para la decoloración de los colorantes remazol azul brillante R (antraquinona), amarillo 145, negro 5, rojo 195 (azoico) y verde malaquita (trifenilmetano). Las concentraciones de nitrógeno no afectaron la producción de lacasa, excepto la mayor concentración (9,6 g/L) que redujo la actividad enzimática. La adición de cobre al medio de cultivo (150 µM) aumentó la actividad de la lacasa en un 112%. La mayor actividad de lacasa (~34300 U/L) promovió la decoloración de los colorantes remazol azul brillante R (67,5%) y verde malaquita (28,3%) en 24h, siendo que los colorantes azoicos fueran parcialmente decolorados. Se concluye que la melaza de soja es un desecho agroindustrial adecuado para la producción de lacasa de P. sanguineus con potencial para degradación de colorantes.

Ethanol induction of laccase depends on nitrogen conditions of Pycnoporus sanguineus

Background Ethanol has been pointed out as a laccase inducer. However, there are controversial reports about its efficiency with some fungi. In this study, we hypothesized that ethanol laccase induced in Pycnoporus sanguineus depends on nitrogen nutriment conditions. To prove this, we assessed laccase production in submerged cultures of P. sanguineus, with different nitrogen concentrations and with, or without ethanol added in a factorial designed experiment. Results In order to analyze the effects of factors on the response variables, a factorial ANOVA, and response-surface models were performed. It was found that the nitrogen source was the main factor that affected laccase production in P. sanguineus. The treatments with yeast extract (2 g/L) and ethanol (3 g/L) induced the highest laccase activity (31.01 ± 4.9 U/L), while the treatments with urea reached the lowest activity (less than 1.6 U/L). Ethanol had positive and synergic effects on laccase production, in accordance with the surface response model, as long as simple nitrogen sources (urea) were not available. Conclusions We suggest that laccase in P. sanguineus is regulated by a catabolic nitrogen repression mechanism; laccase activity is strongly inhibited by urea used as nitrogen source and it decreases when the amount of urea increases; contrarily, a synergic positive effect was observed between yeast extract and ethanol on laccase production.

Atividade "in vitro" de extratos de Pycnoporus sanguineus e Lentinus crinitus sobre o fitopatógeno Fusarium sp / Activity "in vitro" of extracts from Pycnoporus sanguineus and Lentinus crinitus on the pathogen Fusarium sp

The alternative control of plant diseases aims to minimize environmental impacts through the use of natural products. The objective of this work was to evaluate the in vitro activity of aqueous and hydroalcoholic extracts from Pycnoporus sanguineus and Lentinus crinitus against Fusarium sp. A fungus known to cause disease in plants. Fungis amples were collected in the urban and rural areas of Parintins-AM, and tested against different extracts concentrations. Inhibition of mycelia growth, inhibition of conidial germination and inhibition of germination of sclerotia were assessed. Mycelial growth inhibition was highest with hydroalcoholic cold extracts. Hydroalcoholic and aqueous extracts from P. sanguineus obtained by ultrasonic ation and hydroalcoholic cold extract from L. crinitus caused 92% of conidia sporulation. Aqueous hot extracts inhibited sclerotia germination in both fungi samples as well as hydroalcoholic cold extract from L. crinitus. Fungi consortium inhibited sclerotia germination at 1000 µg mL-1 concentration.

O controle alternativo de doenças de plantas tem como objetivo minimizar o impacto ambiental através da utilização de produtos naturais. Assim, este trabalho teve como objetivo avaliar a atividade in vitro de extrato aquoso e hidroalcoólico de Pycnoporus sanguineus e Lentinus crinitus contra Fusarium sp., conhecido por causar doenças das culturas. Os fungos foram coletados em áreas urbanas e rurais de Parintins-AM, e testados em diferentes concentrações de extratos, sendo avaliadas a inibição de crescimento micelial, a inibição da germinação de conídios e a inibição da germinação de esclerócios. Os melhores resultados de inibição do crescimento micelial foram obtidos com extratos hidroalcoólicos frios. Extratos de P. sanguineus obtidos em solvente hidroalcoólico frio e extrato aquoso ultrassônico e extrato de L. crinitus de solvente hidroalcoólico frio, inibiram mais de 92% da esporulação de conídios. Extratos aquosos quentes inibiram a germinação de escleródios, bem como o extrato de P. sanguineus hidroalcoólico frio. O consórcio dos fungos inibiram a germinação de escleródios em 1000 µg mL-1.

Cultivo del hongo Pycnoporus sanguineus (L: Fr) Murr. para evaluar la actividad leishmanicida en promastigotes de leishmania / Cultivation of the fungus Pycnoporus sanguineus (L: Fr) Murr. to evaluate leishmanicidal activity in promastigotes of leishmania

El presente trabajo, tuvo como objetivo el cultivo del hongo Pycnoporus sanguineus (L: Fr) Murr para evaluar la actividad leishmanicida de los extractos de los carpóforos silvestres y cultivados. Los cuerpos fructíferos se colectaron en el distrito de Quellouno de la provincia de la Convención, los cuales fueron trasladados al Centro de Investigación y Producción de Hongos Alimenticios y Medicinales (CIPHAM) donde se realizó el proceso del cultivo, determinando como medio sólido óptimo para la propagación del micelio vegetativo el Agar extracto de malta con una velocidad de crecimiento micelial de 11,7 mm/día; siendo el sustrato óptimo para la obtención de cuerpos fructíferos el que contiene 19 % de salvado de arroz + 80 % de aserrín + 1 % de yeso (SAS), el rendimiento obtenido en este sustrato fue de 41 ,98 %, la eficiencia biológica de 83,97 % con una tasa de producción de 0,23 obtenidos a una temperatura de 16 °C. Los carpóforos silvestres y cultivados fueron llevados al Laboratorio de Química Orgánica, donde se obtuvieron los extractos etanólicos, evaluándose la Biotoxicidad en nauplios de Artemia salina Leach, determinando mediante el Método Probit que la dosis letal media (DL50) para el extracto de los carpóforos silvestres es de 415,04 ug/mL y de los cultivados es 441,20 ug/mL, ambos valores se encuentran en la categoría de extractos moderadamente tóxicos. La actividad leishmanicida de los carpóforos silvestres y cultivados se evaluó en promastigotes de cuatro especies de Leishmania determinándose para cada una de ellas la concentración efectiva media (CEso) así en Leishmania (Viannia) braziliensis 229,58 y 197,76 ug/mL, en Leishmania (Viannia) guyanensis 180,68 y del 172,11 ug/mL, en Leishmania (Viannia) panamensis55, 72 y 37,71 ug/mLy en Leishmania spp.275, 78 y 219,86 ug/mL; respectivamente el análisis estadístico demostró que ambos extractos tienen la misma actividad leishmanicida.

Controle de crestamento bacteriano comum (Xanthomonas axonopodis Ppv. phaseoli) e alterações bioquímicas em feijoeiro induzidas por Pycnoporus sanguineus / Control of bacterial blight (Xanthomonas axonopodis pv. phaseoli) and biochemical analyses of bean resistance treated with Pycnoporus sanguineus extracts

O objetivo deste trabalho foi verificar as atividades antibacteriana e indutora de resistência de extratos de Pycnoporus sanguineus para controle do crestamento bacteriano comum, causado por Xanthomonas axonopodis pv. phaseoli, em feijoeiro. In vitro foram utilizados extratos aquosos de basidiocarpo, micélio e filtrado de cultura de P. sanguineus nas concentrações de 1, 5, 10, 15 e 20%, além das testemunhas água, acibenzolar-S-metil (ASM - 125 mg i.a. L-1) e antibiótico (22,5 mg L-1 de oxitetraciclina + 225 mg L-1 de estreptomicina). In vivo foram realizadas avaliações de severidade e atividade de peroxidase, polifenoloxidase, ß-1,3 glucanase e fenilalanina amônia-liase, com o uso de extrato aquoso de micélio e de basidiocarpo e filtrado de cultura de P. sanguineus a 5% e 10%. Verificou-se atividade antibacteriana apenas para o filtrado de cultura em concentrações acima de 15% e para o extrato de basidiocarpo nas concentrações de 1 a 20%. In vivo, os resultados indicaram o potencial de extratos de basidiocarpos de P. sanguineus para o controle de X. axonopodis pv. phaseoli em feijoeiro, com redução média de 56% na severidade, o que pode ter ocorrido tanto por atividade antimicrobiana direta quanto por indução de resistência, envolvendo principalmente a ativação das enzimas de defesa vegetal peroxidase e polifenoloxidase.

The aim of this work was to verify the antimicrobial and resistance induction activities of Pycnoporus sanguineus extracts for the control of common bacterial blight caused by Xanthomonas axonopodis pv. phaseoli. In vitro assays were performed using aqueous extracts from basidiocarp, mycelium and culture filtrate of P. sanguineus in concentrations of 1, 5, 10, 15 and 20%, with water, acibenzolar-S-methyl (ASM - 125 mg a.i. L-1) and antibiotic (oxytetracycline 22.5 mg L-1 + streptomycin 225 mg L-1) as control treatments. For the in vivo assays the disease severity and the activities of peroxidase, polyphenol oxidase, ß-1,3 glucanase and phenylalanine ammonia-lyase were evaluated using extracts of mycelium, basidiocarp and culture filtrate of P. sanguineus at 5% and 10%. Antibacterial activity was verified only for culture filtrate in concentrations above 15% and for concentrations of 1% to 20% of basidiocarp extract. The results of the in vivo assays indicated the potential of basidiocarps extracts from P. sanguineus for the control of X. axonopodis pv. phaseoli in beans, with an average severity reduction of 56%, which may have been due either to direct antimicrobial activity or to resistance induction involving mainly the activation of the plant defense enzymes peroxidase and polyphenol oxidase.

Degradation of Three Aromatic Dyes by White Rot Fungi and the Production of Ligninolytic Enzymes

This study was conducted to evaluate the degradation of aromatic dyes and the production of ligninolytic enzymes by 10 white rot fungi. The results of this study revealed that Pycnoporus cinnabarinus, Pleurotus pulmonarius, Ganoderma lucidum, Trametes suaveolens, Stereum ostrea and Fomes fomentarius have the ability to efficiently degrade congo red on solid media. However, malachite green inhibited the mycelial growth of these organisms. Therefore, they did not effectively decolorize malachite green on solid media. However, P. cinnabarinus and P. pulmonarius were able to effectively decolorize malachite green on solid media. T. suaveolens and F. rosea decolorized methylene blue more effectively than any of the other fungi evaluated in this study. In liquid culture, G. lucidum, P. cinnabarinus, Naematoloma fasciculare and Pycnoporus coccineus were found to have a greater ability to decolorize congo red. In addition, P. cinnabarinus, G. lucidum and T. suaveolens decolorized methylene blue in liquid media more effectively than any of the other organisms evaluated in this study. Only F. fomentarius was able to decolorize malachite green in liquid media, and its ability to do so was limited. To investigate the production of ligninolytic enzymes in media containing aromatic compounds, fungi were cultured in naphthalene supplemented liquid media. P. coccineus, Coriolus versicolor and P. cinnabarinus were found to produce a large amount of laccase when grown in medium that contained napthalene.