Light conditions affect the growth, chemical composition, antioxidant and antimicrobial activities of the white-rot fungus Lentinus crinitus mycelial biomass.

The mycelial biomass of basidiomycetes is a promising source of compounds and represents an alternative for industrial and biotechnological applications. Fungi use light as information and hold photoresponse mechanisms, in which sensors respond to light wavelengths and regulate various biological processes. Therefore, this study aimed to investigate the effects of blue, green, and red lights on the growth, chemical composition, and antioxidant and antimicrobial activity of Lentinus crinitus mycelial biomass. The chemical composition of the mycelial biomass was determined by chromatographic methods, antioxidant activity was analyzed by in vitro assays, and antimicrobial activity was investigated by the microdilution assay. The highest mycelial biomass yield was observed under blue-light cultivation. Many primordia arose under blue or green light, whereas the stroma was formed under red light. The presence of light altered the primary fungal metabolism, increasing the carbohydrate, tocopherol, fatty acid, and soluble sugar contents, mostly mannitol, and reducing the protein and organic acid concentrations. Cultivation under red light increased the phenol concentration. In contrast, cultivation under blue and green lights decreased phenol concentration. Benzoic and gallic acids were the main phenolic acids in the hydroalcoholic extracts, and the latter acids increased in all cultures under light, especially red light. Mycelial biomass cultivated under red light showed the highest antioxidant activity in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The ferric reducing antioxidant power (FRAP) method showed that all light wavelengths increased the antioxidant activity of mycelial biomass, with the highest value under red light. Moreover, the ß-carotene/linoleic acid co-oxidation (BCLA) assay demonstrated that the antioxidant activity was affected by light cultivation. Mycelial biomass grown under all conditions exhibited antibacterial and antifungal activities. Thus, mycelial biomass cultivation of L. crinitus under light conditions may be a promising strategy for controlling the mycelial chemical composition and biomass yield.

Levantamento epidemiológico dos casos de tuberculose no Brasil e ações alternativas para auxiliar no tratamento / Epidemiological survey of tuberculosis cases in Brazil and alternative actions to aid in treatment / Estudio epidemiológico de los casos de tuberculosis en Brasil y acciones alternativas para ayudar al tratamiento

tuberculose é uma das doenças infectocontagiosas de maior importância no Brasil e no mundo. Afeta de forma importante populações em situação de vulnerabilidade social e econômica. O objetivo deste estudo foi realizar um levantamento do número de casos notificados de tuberculose no Brasil nos últimos 10 anos (2011 a 2021), avaliar os fatores que afetam a transmissão, bem como discutir o tratamento padrão e com fitoterápicos. O levantamento epidemiológico dos casos de tuberculose no Brasil de janeiro de 2011 a dezembro de 2021 foi realizado dentre os notificados pelo Sistema de Informação de Agravos de Notificação (SINAN). Os resultados indicaram um aumento linear de casos a partir de 2017 com 90.776 casos diagnosticados, em 2018 (94.720) e 2019 (96.655). Acredita-se que o aumento linear da tuberculose neste período pode estar relacionado principalmente com o aumento da pobreza, contudo o compartilhamento de utensílios durante o uso de narguilé podem representar fatores de risco para tuberculose. Seis plantas medicinais afetam diretamente as micobactérias (Chenopodium ambrosioides, Tetradenia riparia, Physalis angulata, Origanum vulgare, Eucalyptus globulus, Mikania glomerata) e cinco plantas com atividade antibacteriana auxiliam no trato respiratório (Nasturtium officinale, Allium sativum, Schinus terebinthifolius, Adiantum capillus-veneris, Allium cepa). Contudo, a tuberculose é uma doença reemergente sendo necessária a adoção de políticas públicas que intensifiquem e implementem medidas sócio-educativas para a implantação do uso de fitoterápicos como medida complementar.

Tuberculosis is one of the most important infectious diseases in Brazil and worldwide. It significantly affects populations in situations of social and economic vulnerability. This study aimed to survey the number of reported tuberculosis cases in Brazil in the last 10 years (2011 to 2021) to assess the factors that affect the transmission and discuss standard and herbal treatments. The epidemiological survey of tuberculosis cases in Brazil from January 2011 to December 2021 was carried out among those notified by the Notifiable Diseases Information System (SINAN). The results indicated a linear increase in cases from 2017, with 90,776 diagnosed cases, in 2018 (94,720) and 2019 (96,655). It is believed that the linear increase in tuberculosis in this period may be mainly related to the increase in poverty. However, the sharing of utensils during the use of hookah may represent risk factors for tuberculosis. Six medicinal plants directly affect mycobacteria (Chenopodium ambrosioides, Tetradenia riparia, Physalis angulata, Origanum vulgare, Eucalyptus globulus, Mikania glomerata), and five plants with antibacterial activity help in the respiratory tract (Nasturtium officinale, Allium sativum, Schinus terebinthifolius, Adiantum capillus-veneris, Allium cepa). However, tuberculosis is a re-emerging disease, and it is necessary to adopt public policies that intensify and implement socio-educational measures for using herbal medicines as a complementary measure.

La tuberculosis es una de las enfermedades infecciosas más importantes en Brasil y en el mundo. Afecta significativamente a las poblaciones en situación de vulnerabilidad social y económica. El objetivo de este estudio fue realizar una encuesta sobre el número de casos notificados de tuberculosis en Brasil en los últimos 10 años (2011 a 2021), para evaluar los factores que afectan a la transmisión, así como para discutir el tratamiento estándar y con fitoterapias. La encuesta epidemiológica de los casos de tuberculosis en Brasil desde enero de 2011 hasta diciembre de 2021 se realizó entre los notificados por el Sistema de Informação de Agravos de Notificação (SINAN). Los resultados indicaron un aumento lineal de casos desde 2017 con 90.776 casos diagnosticados, en 2018 (94.720) y 2019 (96.655). Se cree que el aumento lineal de la tuberculosis en este periodo puede estar relacionado principalmente con el aumento de la pobreza, aunque el hecho de compartir los utensilios durante el uso de la shisha puede representar factores de riesgo para la tuberculosis. Seis plantas medicinales afectan directamente a las micobacterias (Chenopodium ambrosioides, Tetradenia riparia, Physalis angulata, Origanum vulgare, Eucalyptus globulus, Mikania glomerata) y cinco plantas con actividad antibacteriana ayudan a las vías respiratorias (Nasturtium officinale, Allium sativum, Schinus terebinthifolius, Adiantum capillus-veneris, Allium cepa). Sin embargo, la tuberculosis es una enfermedad reemergente siendo necesaria la adopción de políticas públicas que intensifiquen e implementen medidas socioeducativas para la implementación del uso de fitoterápicos como medida complementaria.

Estudo do potencial antiviral sobre o Sars-Cov-2 dos compostos identificados no extrato bruto das folhas de Tetradenia riparia / Study of antiviral potential on Sars-Cov-2 of compounds identified in Tetradenia riparia leaves crude extract / Estudio del potencial antiviral sobre el Sars-Cov-2 de los componentes identificados en el extracto crudo de las hojas de Tetradenia riparia

A COVID-19 surgiu de forma repentina, acometendo milhões de pessoas e causando muitas mortes no mundo todo. Diante disso, torna-se necessário a busca de substâncias bioativas com propriedades antivirais. No Brasil, a espécie Tetradenia riparia foi inserida como planta ornamental exótica, com aroma intenso e agradável, sendo cultivada em parques, jardins, residenciais e hortos. O objetivo deste estudo foi identificar compostos presentes no extrato bruto das folhas de Tetradenia riparia com interesse antiviral. O extrato bruto das folhas secas foi obtido por maceração dinâmica por esgotamento do solvente e após, concentrado em evaporador rotativo. A composição química do extrato bruto foi analisada por cromatografia líquida de ultra eficiência acoplada à espectrometria de massas de alta resolução (UHPLC-ESI/qTOF). Foram identificados 31 compostos que foram investigados por meio de levantamento bibliográfico quanto ao seu potencial anti- SARS-CoV-2. Os compostos rosmanol, procianidina, cianidina, betulina, ácido betulínico e o ácido sagerínico, apresentaram potencial atividade antiviral sobre o SARS-CoV-2. Esta investigação é promissora, indicando possivelmente que no extrato bruto das folhas de T. ripária existem compostos que podem combater o SARS-CoV-2. Neste sentido, estudos de ancoramento molecular (docking) e análises in silico sobre a proteína Mpro do vírus devem ser realizadas corroborando desta forma a ação dos compostos identificados.

COVID-19 appeared suddenly, affecting millions of people and causing many deaths worldwide. Therefore, it is necessary to search for bioactive substances with antiviral properties. In Brazil, Tetradenia riparia was inserted as an exotic ornamental plant, with an intense and pleasant aroma, cultivated in parks, residential and vegetable gardens. This study aimed to identify compounds present in the crude extract of Tetradenia riparia leaves with antiviral interest. The crude extract of the dried leaves was obtained by dynamic maceration with solvent exhaustion and then concentrated in a rotary evaporator. The chemical composition of the crude extract was analyzed by ultra- performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC- ESI/qTOF). We identified 31 compounds investigated through a literature review for their anti- SARS-CoV-2 potential. The compounds rosmanol, procyanidin, cyanidin, betulin, betulinic acid, and sagerinic acid showed potential antiviral activity against SARS-CoV-2. Therefore, this investigation is promising, possibly indicating that in the crude extract of T. riparia leaves, there are compounds that can fight SARS-CoV-2. In this sense, molecular docking studies and in silico analyzes on the virus Mpro protein must be carried out, thus corroborating the action of the identified compounds.

SARS-CoV-19 ha aparecido repentinamente, afectando a millones de personas y causando muchas muertes en todo el mundo. Por ello, se hace necesaria la búsqueda de sustancias bioactivas con propiedades antivirales. En Brasil, la especie Tetradenia riparia ha sido introducida como planta ornamental exótica, con un aroma intenso y agradable, siendo cultivada en parques, jardines, residencias y centros de jardinería. El objetivo de este estudio fue identificar los compuestos presentes en el extracto crudo de las hojas de Tetradenia riparia con interés antiviral. El extracto crudo de las hojas secas se obtuvo por maceración dinámica por agotamiento del disolvente y después, se concentró en el evaporador rotatorio. La composición química del extracto crudo se analizó mediante cromatografía líquida de ultra rendimiento acoplada a espectrometría de masas de alto rendimiento (UHPLC-ESI/qTOF). Se identificaron 31 compuestos y se investigó su potencial anti-SARS-CoV-2 mediante un estudio bibliográfico. Los compuestos rosmanol, procianidina, cianidina, betulina, ácido betulínico y ácido sagerínico, mostraron una potencial actividad antiviral sobre el SARS-CoV-2. Esta investigación es prometedora, pues posiblemente indica que en el extracto crudo de las hojas de T. riparia hay compuestos que pueden combatir el SARS-CoV-2. En este sentido, deben realizarse estudios de docking y análisis in silico sobre la proteína Mpro del virus para corroborar la acción de los compuestos identificados.

Lithium bioaccumulation in Lentinus crinitus mycelia grown in media with different lithium sources and pH values.

Lentinus crinitus bioaccumulates lithium in mycelia, but bioaccumulation may be affected by pH of the culture medium. Lithium is used in clinical practice as a mood stabilizer and antidepressant. This study aimed to assess the effect of culture medium pH and lithium source (LiCl or Li2CO3) on lithium bioaccumulation in vegetative mycelia of L. crinitus grown in malt extract broth. Lentinus crinitus U9-1 was cultured in malt extract broth supplemented with Li2CO3 or LiCl (50 mg L-1 lithium) in the pH range of 3.0 to 6.0. The pH was adjusted using HCl solution. The results showed that medium pH affected mycelial biomass production, lithium bioaccumulation in mycelial biomass, and lithium transfer from the culture medium to mycelial biomass. The effect of lithium source on the bioaccumulation capacity of mycelial biomass varied according to pH. At pH 4.0, both lithium sources stimulated mycelial biomass production compared to the control without the addition of lithium. At pH 5.5, Li2CO3 provided the highest lithium bioaccumulation in mycelial biomass. Lithium transfer from the culture medium to mycelia was highest in Li2CO3-supplemented cultures at pH 4.5. LiCl reduced hyphal width compared with Li2CO3 and the control. However, pH and lithium sources did not affect the formation of clamp connections in hyphae. For the first time, the influence of the pH of the culture medium on lithium bioaccumulation by Lentinus crinitus is reported. Finally, we conclude that the culture medium pH affected lithium transfer and bioaccumulation in mycelial biomass differently depending on the lithium source. Additionally, we report the presence of clamp connections in the hyphae of L. crinitus as an indicator of even growth.

Use of green light to improve the production of lignocellulose-decay enzymes by Pleurotus spp. in liquid cultivation.

The influence of green light on mycelium biomass growth and extracellular enzyme activities of edible mushrooms from the Pleurotus genus, which is popularly cultivated all over the world, were investigated. The mycelium of seven strains of five species of Pleurotus (P. citrinopileatus, P. djamor, P. eryngii, P. ostreatus, and P. pulmonarius) was grown in liquid medium at 28 °C in the dark or under green light (515-530 nm). The light source was light-emitting diodes (LED) with photon flux density adjusted to 20 µmol m-2 s-1 that was kept on throughout the cultivation period. After 12 days of growth, the mycelium was recovered and used for biomass determination and the cultivation medium was used to total cellulase, endoglucanase, xylanase, and laccase activities determination. Green light reduced the mycelial biomass growth of Pleurotus spp. but increased the cellulolytic and xylanolytic activities. The cellulolytic activity of most strains increased in the presence of green light with increases ranging from 1.5 times (P. ostreatus endoglucanase) to 8 times (P. citrinopileatus total cellulase and endoglucanase). Green light reduced laccase activity for most strains with the greatest reduction for P. eryngii (2.2 times lower). The specific enzymatic activity of cellulase and endoglucanase from P. citrinopileatus, increased by 31 times and 30 times, respectively, compared to the dark. Also, the specific laccase and xylanase activities of P. pulmonarius increased 4.4 times and 6.8 times, respectively, under green light. The use of light at particular wavelengths can be a viable strategy to increase the production of enzymes for different biotechnological applications and species of Pleurotus are particularly interesting for this purpose.

Lentinus crinitus response to blue light on carbohydrate-active enzymes / Resposta de Lentinus crinitus à luz azul em enzimas ativas por carboidratos

Fungi are capable of sensing light from ultraviolet to far-red and they use light as a source of information about the environment anticipating stress and adverse conditions. Lentinus crinitus is a lignin-degrading fungus which produces laccase and other enzymes of biotechnological interest. The effect of blue light on fungal enzymatic activity has been studied; however, it has not been found studies on the effect of the blue light on carbohydrate-active enzymes and on mycelial biomass production of L. crinitus. We aimed to investigate carbohydrate-active enzymes activity and mycelial biomass production of L. crinitus cultivated under continuous illumination with blue light. L. crinitus was cultivated in malt extract medium in the dark, without agitation, and under continuous illumination with blue light-emitting diodes. The blue light reduced the total cellulase, pectinase and xylanase activities but increased the endoglucanase activity. Blue light reduced the mycelial growth of L. crinitus in 26% and the enzymatic activity-to-mycelial biomass ratio (U mg-1 dry basis) increased in 10% total cellulase, 33% endoglucanase, and 16% pectinase activities. Also, it is suggested that L. crinitus has a photosensory system and it could lead to new process of obtaining enzymes of biotechnological interest.

Fungos são capazes de sentir a luz com comprimentos de onda que variam do ultravioleta ao infravermelho e usam a luz como fonte de informação sobre o ambiente, antecipando condições adversas e de estresse. Lentinus crinitus é um fungo ligninolítico que produz lacase e outras enzimas de interesse biotecnológico. O efeito da luz azul na atividade enzimática de fungos já foi estudado, contudo, ainda não há estudos sobre o efeito da luz azul na produção de enzimas ativas sobre carboidratos (CAZymes, carbohydrate-active enzymes) e de biomassa micelial de L. crinitus. O objetivo deste estudo foi investigar a avitivade de CAZymes e a produção de biomassa micelial de L. crinitus cultivado sob iluminação continua com luz azul. L. crinitus foi cultivado em meio extrato de malte, sem agitação, na ausência de luz e sob luz continua fornecida por diodos emissores de luz azul. A luz azul reduziu a atividade de cellulase total, pectinase e xilanase, mas aumentou a atividade de endoglucanase. A luz azul reduziu o crescimento micelial de L. crinitus em 26% e aumentou a razão atividade enzimática/biomassa micelial (U mg-1 em base seca) de cellulase total em 10%, endoglucanase em 33% e pectinase em 16%. Além disso, sugere-se que L. crinitus possua um sistema fotossensorial que poderia ser explorado para a otimização de bioprocessos que visam a obtenção de enzimas de interesse biotecnológico.

Decolorization of azo and anthraquinone dyes by crude laccase produced by Lentinus crinitus in solid state cultivation.

White-rot basidiomycetes such as Lentinus crinitus produce laccases with potential use in dye biodegradation. However, high productivity and enzymes with specific properties are required in order to make viable laccase production. We aimed to produce laccase from Lentinus crinitus grown in sugarcane bagasse for dye decolorization. Solid state cultivation medium had sugarcane bagasse added with a nutrient solution of 10 g/L glucose, 1 g/L KH2PO4, 0.5 g/L MgSO4, 0.001 g/L FeSO4, 0.01 g/L ZnSO4, and 0.01 g/L MnSO4. The addition of different nitrogen sources (peptone, urea, or peptone plus urea) and different nitrogen concentrations (0, 0.4, 0.6, 0.8, 1.0, and 1.2 g/L) were evaluated. Enzymatic extract was used in the decolorization of azo dyes, reactive blue 220 (RB220) and reactive black 5 (RB5), and anthraquinone dye, Remazol brilliant blue R (RBBR). The greatest laccase activity (4800 U/g dry mass) occurred when the peptone and urea mixture was added to the solid state cultivation medium. When the nitrogen concentration was 1 g/L, the laccase activity increased to 6555 U/g dry mass. The laccase activity peak occurred on the 10th day, and the maximum decolorization within 24 h was observed with enzymatic extracts obtained on different cultivation days, i.e., 6th day for RB220, 10th day for RB5, and 9th day for RBBR. Manganese and lignin peroxidases were not produced when nitrogen was added to the cultivation medium. The crude enzymatic extract was more effective in the decolorization of azo dyes (RB220 and RB5), more than 90% of decolorization, than anthraquinone dye with 77% decolorization.

Lithium bioaccumulation in Lentinus crinitus mycelial biomass as a potential functional food.

Lentinus crinitus is an important basidiomycete consumed by ethnic groups from the Amazon, commonly found in decomposing trees with high lignolytic and antioxidant activities. Lithium is a mood stabilizer, antiepileptic, antipsychotic, and antidepressant used in clinical practice. This study aimed to evaluate L. crinitus mycelial biomass bioaccumulated with lithium in liquid cultivation medium. The malt extract medium was added from zero to 100 mg L-1 lithium from two lithium sources (Li2CO3 and LiCl). The maximum mycelial biomass production was 7218.89 mg L-1 in the culture medium added with 5 mg L-1 lithium from LiCl. The highest lithium concentration in the mycelial biomass was of 574.72 µg g-1 produced in the culture medium with 25 mg L-1 lithium from Li2CO3. Pearson's correlation showed that Li2CO3 reduces the mycelial biomass and increases lithium bioaccumulation. The maximum translocated lithium from cultivation medium to mycelial biomass was up to 19 or 28% with LiCl or Li2CO3, respectively. Therefore, although Li2CO3 presents greater inhibition on the mycelial biomass production, it promoted greater lithium bioaccumulation in L. crinitus mycelial biomass and resulted in greater yield of lithium translocation. The equivalent daily dose of lithium for psychiatric treatment, without bioavailability studies, could be reached with 97.4 g lithium-enriched mycelial biomass and, based in the literature, for reduction of violence and criminality rates the amount could be reached with 0.24-0.58 mg. Thus, the development of lithium-enriched mycelial biomass could be an alternative functional food.

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.