Trametes versicolor (L.) Lloyd as a source of thermostable serine protease: production and characterization

Proteases are ubiquitously present and are among the largest groups of commercially important enzymes. Here, we investigated a wood-rot basidiomycete Trametes versicolor (L.) Lloyd [Syn. Coriolus versicolor (L.) Quél.; Polyporus versicolor (L.) Fr.] as a source of the enzyme serine protease, its production, and optimized to obtain a higher yield of the enzyme.. The significant variables with optimized values for maximum production of the enzyme were temperature (30?C), incubation time (120 h) and wheat bran (10 g). The yield increased by 30.76% by statistically optimizing the media. The optimized temperature and pH for the maximum protease activity was 50?C and pH 7.0, respectively. The enzyme was purified through ion exchange (using DEAE cellulose 52 resin) and gel filtration chromatography (using Superdex 200 column). The purified enzyme had a retention time of 7 min in RP-HPLC. The enzyme was stable at a broad range of temperature (30-60?C) and pH (5.0-8.0) with a half-life of 58.72 min, Vmax of 37.17 ?M min/mL and Km of 0.657 mg/mL. Its activity was enhanced by Na+, Ca2+, Mg2+ ions and SDS surfactant. These properties make this enzyme a valuable candidate for industrial applications

Polissacarídeos de Lentinula edodes suprimem a expressão de citocinas pró-inflamatórias e de colite em camundongos / Lentinula edodes polysaccharides suppressed pro-inflammatory cytokines expression and colitis in mice

ABSTRACT Background: Polysaccharides from edible mushrooms possess immunomodulatory, anti-inflammatory, and anti-tumor activities. Recent studies indicated that necroptosis plays a role in the pathogenesis of inflammatory diseases and mediates increased expression of inflammatory cytokines. Objective: Therefore, it is imperative to determine the impact of polysaccharide extract from Lentinula edodes (L. edodes) on inflammatory cytokines in experimental model of colitis in mice. Methods: Female C57BL/6 mice divided into three or four mice per group were used for this study. Polysaccharide sample was orally administered to mice prior to (7 days) and during colitis induction with 2.5% dextran sodium sulfate (7 days), followed by additional 3 days of administration. Changes in body weight and colon length were used as markers for colitis, and pro-inflammatory cytokines and tumor necrosis factor receptor 1 (TNFR1) expressions, as well as necroptosis were analyzed in the colon of colitis mice. Data obtained were analysed by Tukey-Kramer and two-tailed standard t tests. Results: The results indicated that the polysaccharide sample suppressed colitis in mice using effects on the body weight and colon length as markers. Also, it was demonstrated that necrostatin-1, a specific inhibitor of necroptosis, suppressed the expression of interleukin (IL)-8, a pro-inflammatory chemokine, in Caco-2 cells induced necroptosis induced by zVAD and TNF-α, an indication that necroptosis may be involved in the expression of pro-inflammatory cytokines. Moreover, the polysaccharide sample suppressed the expression of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-α, IL-6, IL-1β, and interferon (IFN)-γ in the colon of mice. Conclusion: These results suggested that the suppressive effects of the polysaccharide sample on inflammatory cytokines expression may contribute to its anti-colitis effect, and so may serve as a potent therapeutic agent against inflammatory bowel disease.

RESUMO Contexto: Polissacarídeos de cogumelos comestíveis possuem atividades imunomodulatórias, anti-inflamatórias e anti-tumorais. Estudos recentes indicaram que a necroptose desempenha um papel na patogênese de doenças inflamatórias e regula o aumento da expressão de citocinas inflamatórias. Objetivo: Torna-se imprescindível determinar o impacto do extrato de polissacarídeo de Lentinula edodes (L. edodes) em citocinas inflamatórias em modelo experimental de colite em camundongos. Métodos: Foram utilizados para este estudo os camundongos C57BL/6 femininos divididos em três ou quatro camundongos por grupo. A amostra de polissacarídeo foi administrada oralmente em camundongos antes (7 dias) e durante a indução de colite com sulfato de dextran sulfato de sódio (7 dias), seguido por 3 dias adicionais de administração. Alterações no peso corporal e comprimento do cólon foram utilizadas como marcadores para colite, e citocinas pró-inflamatórias e tumores receptor fator 1 (TNFR1), bem como necroptose foram analisadas no cólon de camundongos colite. Os dados obtidos foram analisados por testes Tukey-Kramer e testes padrão t de duas caudas. Resultados: Os resultados indicaram que a amostra de polissacarídeo suprimiu colite em camundongos usando efeitos sobre o peso corporal e o comprimento do cólon como marcadores. Além disso, foi demonstrado que a necrostatina-1, inibidora específica da necroptose, suprimiu a expres são de interleucina (IL)-8, uma quimiocina pró-inflamatória, em células caco-2 induzidas necroptose induzidas por zVAD e TNF-α, uma indicação de que a necroptose pode estar envolvida na expressão de citocinas pro-inflamatórias. Além disso, a amostra de polissacarídeo suprimiu a expressão de citocinas pró-inflamatórias, como o fator de necrose tumoral (TNF)-α, IL-6, IL-1β e interferon (IFN)-γ no cólon dos camundongos. Conclusão: Esses resultados sugeriram que os efeitos supressivos da amostra de polissacarídeo na expressão de citocinas inflamatórias podem contribuir para o seu efeito anti-colite, podendo, portanto, servir como um potente agente terapêutico contra doença inflamatória intestinal.

Antimicrobial activity of the edible mushroom Pleurotus eryngii(DC. ) Quél grown in liquid medium

Edible mushrooms have a number of medicinal properties and this study aimed to investigate the antimicrobial activity of Pleurotus eryngiiDPUA1816 in metabolic broths after being grown in submerged cultivation. Mycelial fragments of pure P. eryngiiculture was inoculated in sweet potato culture medium and incubated at 150 rpm for 15 days at 25°C. Pleurotuseryngiiwas also cultivated for 18 days under the same conditions, the mycelial biomass was separated by filtration for quantification. The supernatant was used in the diffusion test in agar and performed against Escherichia coliCCCD-E005, Staphylococcus aureus CCCD-S009, Pseudomonas aeruginosaCCCD-P004, Candida albicansCCCD-CC001, Candida parapsilosis CCCD-CC004 and Candida tropicalisCCCD-CC002. The samples showed no inhibitory activity against bacteria, however they showed some activity againstC. albicans(12.17 mm), C. parapsilosis(27.67 mm) and C. tropicalis(13.67 mm). After being cultivated for 18 days, P. eryngiiwas able to inhibit all yeasts after 12 days of culture, with an inhibition halo of 29.33 mm at 16 days against C. parapsilosis. This study demonstrates the antifungal potential filtered liquids from P.eryngiicultivated in purple-skinned sweet potato culture medium, which suggests the possibility of the use of this species by the pharmaceutical industry as a natural source of biological action.

Tecnologic Development on Pleurotus Cultivation: Specific Practices Used in Brazil

HIGHLIGHTS P. ostreatus and P. sapidus are the most productive species under the evaluated conditions. Different growing systems are suitable for the production of P. ostreatus var. Florida. Temperature control level affects differently the P. ostreatus var. Florida isolates. Environmental and strain factors affect yield and production parameters of P. ostreatus var. Florida.

Abstract In Brazil, Pleurotus is the most important mushroom produced especially P. ostreatus var. Florida. In this country as in many others, the great potential for mushroom cultivation remains unexplored. Therefore, it is very important to develop new studies that allow optimizing its production. The aims of the manuscript were: i) to evaluate the productivity of six different species of Pleurotus (P. citrinopileatus; P. djamor; P. ostreatus; P. ostreatus var. Florida; P. pulmonarius; P. sapidus); ii) to measure the effect of three different environmental conditions during cultivation of three isolates of P. ostreatus var. Florida. As results, P. ostreatus and P. sapidus were the most productive isolates under the evaluated conditions. Different environments produced variable effects according to the P. ostreatus var. Florida isolates, being possible to observe a highly plastic strain (POF 02/18), a highly sensitive strain (POF 03/18) and a strain with variable responses (POF 01/18).

Actividad de las enzimas hidrolíticas en cepas del hongo shiitake (Lentinula edodes) cultivadas en pulpa de café / Hydrolytic enzyme activities in shiitake mushroom (Lentinula edodes) strains cultivated on coffee pulp

Se estudió la producción de enzimas hidrolíticas (celulasas, laminarinasas y xilanasas) en cultivos de Lentinula edodes en pulpa de café estéril. Se tomaron muestras de sustrato colonizado por el micelio después de 7, 14, 21, 28 y 35 días de incubación a 25°C (W1 a W5) y durante el período de fructificación en diferentes etapas: formación de primordios (PF), primera cosecha (H) y una semana después de la primera cosecha (PH). La actividad enzimática fue menor al inicio del crecimiento micelial y mostró mayores niveles en la formación y el desarrollo de basidiomas. Durante la etapa reproductiva del hongo, las muestras se sometieron a un tratamiento de remojo. Sin embargo, no fue posible relacionar este tratamiento con el aumento de la producción de enzimas. Los niveles de actividad enzimática sugieren que la secreción de las enzimas estudiadas no influye en la capacidad de adaptación de las cepas al sustrato

Hydrolytic enzyme production (cellulases, laminarinases and xylanases) was studied in cultures of Lentinula edodes on sterilized coffee pulp. Samples of substrate colonized by mycelia were taken after 7, 14, 21, 28 and 35 days of incubation at 25°C (W1 to W5) and during the fruiting period at different stages: formation of primordia (PF), first harvest (H) and one week after the first harvest (PH). The enzymatic activity was lower during the early mycelial growth and showed higher levels during the formation and development of fruiting bodies. During the reproductive stage of the fungus, the samples were subjected to a soaking treatment; however, it was not possible to relate this soaking treatment to the increase in enzyme production. The levels of enzymatic activity suggest that secretion of the studied enzymes does not influence the adaptability of the strains to the substrate

Lentinula edodes based GIS mapping, biometabolites and antiinflamatory activity of wild edible mushrooms from tropical 'sacred grove' forests of Meghalaya, India / Mapas basados en GIS deLentinula edodes, biometabolitos y actividad antiinflamatoria de los hongos silvestres comestibles de los bosques tropicales "bosque sagrado" de Meghalaya, India

AbstractThe biodiversity rich state of Meghalaya, India located in the realms of mega-biodiversity hotspots, is home to numerous species of wild edible macrofungi that are used extensively by the mycophillic ethnic population, as a part of their traditional cuisine and medicine systems. However, habitat loss, due to deforestation and climate change, is destroying the natural population of these mushrooms, depleting their availability to the local communities. In the present investigation, a GIS guided habitat search, using Lentinula edodes as a representative species, was used in mapping the habitats of wild edible macrofungi of the study region. Sampling of around 4 000 specimens per distinct morphological type available in the traditional markets and "sacred grove" forests indicated presence of ten common genera, belonging to nine different families of wild edible mushrooms. Nutritional profiling of the representative species Lentinula edodes was carried out by evaluation of its moisture, total fat, crude protein and carbohydrates contents by standard methods. Similarly, bioactive components determination was performed by estimation of total phenols, flavonoids, ascorbic acid, β-carotene and lycopenes. Bioactivity of the mushrooms extracts was studied using the DPPH radical scavenging and Human Red Blood Cell (HRBC) membrane stabilization assays. The present investigation successfully attempted to explore remote sensing technologies and GIS (Geographic Information System) based system to predict the natural habitats of wild edible mushrooms of Meghalaya, India which we believe will lead to the generation of a mushroom specific non-wood forest resource mapping system in the near future. Results of nutritional profiling and biological activity studies on the representative species of wild edible mushrooms from the studied region revealed that it is a rich source of essential nutrients and antioxidants.

ResumenLa rica biodiversidad del estado de Meghalaya, India situado en los reinos de los hotspots mega-biodiversidad es el hogar de numerosas especies de macro hongos silvestres comestibles que se utilizan ampliamente por la población étnica micofílica como parte de su alimentación y medicina tradicional. Sin embargo, la pérdida de hábitat debido a la deforestación y el cambio climático está destruyendo la población natural de estos hongos, que agotan su disponibilidad para las poblaciones locales. En la presente investigación, se utilizó el SIG para la búsqueda guiada de hábitat usando Lentinula edodes como especie representativa en la cartografía de los hábitats de macrohongos silvestres comestibles de la región de estudio. Alrededor de cuatro mil ejemplares de distinto tipo morfológico fueron muestreados según su disponibilidad en los mercados tradicionales y los "sacred grove" (bosques sagrados) lo que indicó la presencia de diez géneros comunes pertenecientes a nueve familias de setas silvestres comestibles. Perfiles nutricionales de las especies representativas Lentinula edodes se llevaron a cabo mediante la evaluación de su humedad, grasa total, proteína cruda y contenido de carbohidratos. Del mismo modo, la determinación de componentes bioactivos se realizó por estimación de fenoles totales, flavonoides, ácido ascórbico, β-caroteno y licopeno. La bioactividad de los extractos de setas se estudió mediante la captación de radicales DPPH y el ensayo de estabilización de la membrana de glóbulos rojos de la sangre humana (hRBC). Los resultados de estudios de perfiles nutricionales y actividad biológica en hongos comestibles revelan que es una fuente rica en nutrientes esenciales y antioxidantes. La presente investigación ha intentado con éxito explorar las tecnologías de teledetección y SIG (Sistema de Información Geográfica) para predecir el hábitat natural de los hongos silvestres comestibles de Meghalaya, India que creemos dará lugar a la generación de un mapeo de los recursos forestales no madereros, específicamente de hongos, en el futuro cercano.

Extraction, Purification and Characterization of Endo-Acting Pullulanase Type I from White Edible Mushrooms.

Pullulanase (EC 3.2.1.41) has been isolated and purified from white edible mushrooms by ammonium sulphate precipitation (20-70%) followed by ion exchange chromatography (DEAE-cellulose) and gel filtration (Sephadex G 75-120), with final yield (20%) and purification fold (17.8). The molecular mass of pullulanase enzyme was 112 kDa as estimated by SDS-PAGE and the pI value was 6.2. The apparent Km and Vmax values for purified pullulanse on pulluan were 0.27 mM and 0.74 μM min-1 respectively. The activity was optimum at 40○C and pH 6. Pullulanase showed moderate thermo-stability. A relative substrate specificity for hydrolysis of soluble starch, amylopectin and glycogen was 80, 60 and 30% respectively. Enzyme activity was highly activated by Fe+2, Mn+2 and Ca+2 ions, while the activity was inhibited by Hg+2 and Ag+ ions. Ethylenediaminetetraacetic acid (EDTA) and Dithiothreitol (DTT) were activated the enzyme activity. On contarary iodoacetate and sodium fluoride were inhibited the activity. HPTLC (High Performance Thin Layer Chromatography) plate showed that the purified pullulanase caused the complete hydrolysis of pullulan to maltotriose.

Caracterización in vitro y producción de inóculo de cepas guatemaltecas de lepista nuda (Bull.: Fr. ) cooke / Characterization and spawn production of guatemalan strains of lepista nuda (Bull.: Fr. ) cooke

Lepista nuda es un hongo comestible muy apreciado por su delicado sabor, especialmente en varios países de Europa. En Guatemala, lo consumen personas de las etnias Kaqchikel y Mam, por lo que constituye un alimento ancestral que puede aprovecharse como una alternativa alimenticia y económica. Por tal razón, en este estudio se investigaron cinco cepas nativas de L. nuda y se evaluó su crecimiento in vitro en tres medios de cultivo (agar extracto de malta –AEM, agar papa dextrosa -APD- y agar Sabouraud -SAB-) a dos temperaturas (18 y 26°C), así como la producción de inóculo en granos de trigo, cebada y sorgo. Se determinó que el medio más adecuado para el crecimiento micelial de las cepas fue AEM incubado a 26°C. Todas las cepas presentaron colonias de color blanco a lila, textura algodonosa, bordes irregulares, con hifas de 1 a 4 μm y fíbulas en regular cantidad a abundantes en todos los medios evaluados. En la producción de inóculo, las cepas presentaron mayor velocidad de colonización en los granos de trigo. Se recomienda utilizar AEM para la producción de biomasa y granos de trigo para la producción de inóculo de las cepas evaluadas.

Lepista nuda is an edible mushroom highly appreciated for its delicate flavor and in several European countries. In Guatemala is used by people of Kaqchikel and Mam ethnic groups and constituting an ancient food that can be used for economic and nutritional alternatives for the country. For this reason, five native strains of L. nuda were studied in vitro in three culture media (malt extract agar -AEM-, potato dextrose agar -APD- and Sabouraud agar -SAB-) at two different temperatures (18 and 26°C). The spawn production where evaluated in wheat, barley and sorghum grains. The most appropriate medium for the mycelial growth of the strains was AEM incubated at 26°C. All strains showed colonies white to lilac, cottony texture, irregular edges, with hyphae of 1 to 4 μm, and moderate to abundant quantity of clamp connections in all media tested. In the spawn production, the strains showed faster colonization in wheat grains. It is recommend the medium AEM for the production of biomass and wheat grain for spawn production for the strains tested.

Actividad antioxidante de extractos de diez basidiomicetos comestibles en Guatemala / Antioxidant activity of extracts of ten edible basidiomycetes in Guatemala

Los antioxidantes son esenciales en el cuerpo humano para prevenir el daño oxidativo. Estas sustancias pueden obtenerse de diferentes fuentes como frutas, plantas y hongos. En Guatemala, diversas especies de hongos comestibles son comercializadas y consumidas, sin embargo su actividad antioxidante no ha sido documentada en el país. El objetivo de este estudio fue determinar la actividad antioxidante de extractos acuosos y etanólicos obtenidos de diez especies de basidiomicetos comestibles (Agaricus aff. bisporus, Agaricus brunnescens, Armilla-riella polymyces, Amanita garabitoana, Boletus edulis, Cantharellus lateritius, Laccaria amethystina, Lactarius deliciosus, Neolentinus ponderosus y Pleurotus ostreatus). Se utilizó un método cualitativo por cromatografía en capa fina (CCF) y tres ensayos macrométricos in vitro de cuantificación de fenoles totales, reducción del radical 1,1-difenil-2-pricrilhidrazilo (DPPH) y decoloración del radical catiónico del reactivo ácido 2,2’-azinobis-(acido-3-etilbenzotiazolina-6-sulfónico)(ABTS). Los extractos acuosos mostraron mayor actividad antioxidante que los extractos etanólicos en todas las técnicas cuantitativas realizadas. La especie que mostró mayor actividad antioxidante en ambos extractos fue B. edulis, cuyos resultados en el extracto acuoso fueron: fenoles totales 93.46 ± 18.17 mg/g, DPPH CI 50 0.93 mg/mL (IC95% 0.65-1.28) y en ABTS CI50 0.96 mg/mL (IC 95% 0.63-1.35); los resultados en el extracto etanólico fueron: Fenoles totales 42.70 ± 3.48 mg/g, DPPH CI50 2.75 mg/mL (IC 95% 2.46-3.07) y 4.13 mg/mL (IC 95% 2.67-5.88). Se evidencia de esta forma que las especies de basidiomicetos estudiadas presentan actividad antioxidante por lo cual pueden ser una fuente potencial de antioxidantes naturales.

Antioxidants are essential in the body to prevent oxidative damage. These antioxidant substances are obtained from different sources such as fruits, plants and mushrooms. In Guatemala, diverse species of mushrooms are commercialized and consumed, however their antioxidant activity has not been documented in Guatemala. The goal of this study aimed to determine the antioxidant activity of aqueous and ethanolic extracts obtained from edible basidiomycete’s species: Agaricus aff. bisporus, A. brunnescens, Armillariella polymyces, Amanita garabitoana, Boletus edulis, Cantharellus lateritius, Laccaria amethystina, Lactarius deliciosus, Neolentinus ponderosus and Pleurotus ostreatus. Thin layer chromatography (TLC) was used as a qualitative method to determine the presence of antioxidant activity, subsequently, three in vitro macrometric assays were used: the quantification of total phenolics, reduction of 1,1-diphenyl-2-pricrilhidrazil (DPPH) radical, and discoloration of the acid reagent 2.2 ‘-azinobis-(acid-3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cation. Aqueous extracts exhibited higher antioxidant activity than ethanolic in all the quantitative techniques used. The specie that showed greater antioxidant activity in both extracts was B. edulis, whose results of aqueous extract were as follows: Total phenolics 93.46 ± 18.17 mg/g , DPPH IC50 0.93 mg/mL (CI95% 0.65-1.28) and ABTS IC50 0.96 mg/mL (CI95% 0.63-1.35); results of ethanolic extract were: Total phenolics 42.70 ± 3.48 mg/g, DPPH IC50 2.75 mg/mL (CI95% 2.46-3.07) and ABTS IC50 4.13 mg/mL (CI95% 2.67-5.88). Based in the result of the study all the basidiomycete’s species that were evaluated have antioxidant activity, therefore, are potential sources of natural antioxidants.

Evaluación de la producción de cuerpos fructíferos de cepas guatemaltecas del hongo comestible Rukoxil Tunay Che’ (Agrocybe cylindracea (DC. Maire.) en diferentes sustratos / Production of fruiting bodies of Guatemalans strains of the edible mushroom Rukoxil Tunay Che' (Agrocybe cylindracea (DC.) Maire. ) on different substrates

En el presente estudio se determinó la producción de cuerpos fructíferos de cinco cepas nativas de Agrocybe cylindracea sobre tres sustratos y dos tratamientos térmicos, a través del porcentaje de eficiencia biológica y la medición del diámetro de los píleos. Se encontró que el mayor porcentaje de eficiencia biológica de las cepas en los sustratos evaluados fue 115.84 %, que correspondió al sustrato constituido por 29% paja de trigo más 1% de harina de soya pasteurizado y obtenido por la cepa 58.01, la cual también produjo las mayores eficiencias biológicas en todos los sustratos evaluados. Al confrontar el porcentaje de eficiencia biológica en los diferentes sustratos, todas las cepas presentaron valores altos en el sustrato compuesto por 28% de paja de trigo, 1% harina de soya y CaCO3. Con respecto al diámetro de los cuerpos fructíferos, las cepas 58.01, 59.01, 60.01 y 638.08 produjeron píleos menores de 2 cm, entre 2-4 cm y mayores a 4.0 cm en los diferentes sustratos y tratamientos, excepto la cepa 59.01 que en el sustrato formulado con 29% de paja de trigo y 1% de harina de soya, solo produjo cuerpos fructíferos con píleos menores a 2 cm y entre 2-4 cm. En el análisis proximal de los basidiomas de las cepas evaluadas se obtuvo un alto porcentaje de proteínas, fibra cruda y carbohidratos, así como bajo porcentaje de grasas. Se recomienda que en futuras investigaciones o transferencia de tecnología a comunidades o entidades interesadas en el cultivo de este hongo, utilizar paja de trigo suplementada con harina de soya y como regulador de pH CaCO3, ya que en dicho sustrato se obtuvieron los mayores porcentajes de eficiencia biológica para la producción de cuerpos fructíferos de A. cylindracea.

This study determined the production of fruiting bodies of five native strains of Agrocybe cylindracea over three different substrates and two heat treatments, by the biological efficiency percentage and the measurement of diameters of pileus. The major percentage of biological efficiency found from the strains in the evaluated substrates was 115.84%, and corresponded to the substrate formulated by 29% of wheat straw and 1% of pasteurized soy flour, and obtained from the strain 58.01, which also produced the major biological efficiencies in all of the evaluated substrates. When confronting the percentage of biological efficiency in the different substrates, all the strains presented high values in the substrate comprising 28% of wheat straw, 1% of soy flour and CaCO3. In relation to the diameter of the fruiting bodies, the strains 58.01, 59.01, 60.01 and 638.08 produced pileus less than 2 cm, between 2-4 cm and greater than 4 cm in the different substrates and treatments, excepting the strain 59.01 which in the substrate formulated with 29% of wheat straw and 1% of soy flour, only produced fruiting bodies with pileus less than 2 cm and between 2-4 cm. In the chemical proximate analysis of the fruiting bodies of the tested strains, a high percentage of proteins, crude fiber and carbohydrates was obtained, and also a low percentage of fats. For future research or when transferring the technology to communities interested in mushroom cultivation, the utilization of wheat straw supplemented with soy flour and as a regulator of pH CaCO3 is recommended, as in this substrate the highest percentages of biological efficiency for the production of fruiting bodies of A. cylindracea where obtained.

Immunomodulatory Effects of Aqueous Extracts of Auricularia sp and Pleurotus sp Mushrooms in Cyclophosphamide- Immunosuppressed Wistar Rats.

Aims: To determine the immunomodulatory effect of aqueous extracts of Auricularia sp and Pleurotus sp mushrooms using an immunosuppression animal model. Study Design: Pre-clinical experimental study. Place and Duration of Study: Department of Pharmacology and Therapeutics, College of Health Sciences and Division of Pharmacology, Department of Physiological Sciences, School of Veterinary Medicine, Makerere University, between August 2010 and December 2011. Methodology: A total of 80 Wistar rats divided into 8 groups (n=10) were used in the experimental study. Cyclophosphamide (10mg/kg) was administered orally (p.o) to fifty (50) Wistar rats in the first 5 groups for 28 days. In addition, rats in Group I received distilled water, groups II & III received 300mg/kg & 600mgkg of Auricularia sp extract respectively and Groups IV &V received 400mg/kg & 800mg/kg Pleurotus sp extract respectively. Wistar rats in Group VI received only 300mg/kg Auricularia sp extract, group VII received 400mg/kg Pleurotus sp extract and Group VIII received only distilled water. Blood samples were collected on days 0, 14 and 28 to determine the total and differential WBC counts. Data is presented as mean±SEM and analyzed using one-way ANOVA followed by a student’s t-test for statistical significance. Mean values are compared with initial values and the control group (Group VIII). Results: No mortality of Wistar rats was observed over the 28-day experimental period. Cyclophosphamide though caused statistically significant (p<0.05) reduction in total WBC on day 14 and 28 compared with day 0 in control group from 11.26±0.59 on day 0 to 6.11±0.41 day 14, & 4.12±0.22 on day 28. Lymphocytes and Neutrophil counts were also significantly reduced in control group by day 28 compared to mushroom extract treated rats. Results show that aqueous extracts of Auricularia sp & Pleurotus sp mushrooms significantly (p<0.05) moderated the reductions in total & differential WBC on day 14 and 28 as compared to the control group. The mushroom extracts also increased total and differential WBC in normal rats as compared to the normal group (Group VIII). Conclusion: Aqueous extracts of Auricularia sp and Pleurotus sp mushrooms moderated cyclophosphamide-induced reduction in WBC in Wistar rats indicating potential benefit in chemotherapy induced immunosuppression. Application of these mushrooms in immune suppression research appears to be new as reflected in the literature. These are however preliminary data to be more completely documented by further experiments, possibly investigating also some aspect of immune cell functions (e.g. cytotoxicity or cytokine production).

Crescimento micelial de Lentinus sajor caju (Fr.) Fr. E Pleurotus spp. em diferentes resíduos agrícolas / Mycelial growth of Lentinus sajor caju (Fr.) Fr. and Pleurotus spp. in different agricutural wastes

O objetivo deste trabalho foi avaliar o desenvolvimento in vitro de três linhagens de cogumelos [Pleurotus ostreatoroseus Singer, Pleurotus pulmonarius (Fr.) Quél. e Lentinus sajor caju (Fr.) Fr.], reativados em meio de cultura BDA e cultivadas em meios de cultura formulados à base de resíduos agrícolas, como a palha de arroz, casca de mamona, casca de amendoim. No meio sólido formulado foi adicionado um disco de cultura no centro da placa, estas foram incubadas a 25oC até a colonização do meio. Avaliou-se, diariamente, o diâmetro da colônia e obteve-se, aos cinco dias de cultivo, a massa miceliana, sendo os resultados obtidos submetidos a análise de variância e teste de Tukey. O meio contendo extrato da casca de amendoim foi o mais adequado para o crescimento da linhagem utilizada de Lentinus sajorcaju, que colonizou 68,30% da placa de Petri. O meio contendo extrato de casca de mamona foi mais favorável ao crescimento de Pleurotus ostreatoreoseus que colonizou 40,01% da placa. A linhagem nativa de Pleurotus pulmonarius cresceu indiferentemente nos meios testados e teve maior crescimento comparada a linhagem de Pleurotus ostreatoroseus. A linhagem de Lentinus sajor-caju no meio contendo extrato da casca de amendoim, apresentou massa micelial seca significativamente maior que todas as demais. O crescimento micelial das linhagens de Lentinus sajor-caju e Pleurotus ostreatoroseus é influenciado pelo meio de cultivo, enquanto o da linhagem nativa utilizada não é influenciado pelos meios testados.

The aim of this study was evaluate the in vitro development of three mushrooms strains [Pleurotus ostreatoroseus Singer, Pleurotus pulmonarius (Fr.) Quél. e Lentinus sajor-caju (Fr.) Fr.], growing media based in agricultural wastes, such rice straw, castor bean seed husks and peanut shells. In the solid medium was added a culture disc in the center of a petri dish and than was incubated at 25oC even the completly colonization of the medium, the data obtained were submitted to variance analysis and Tukey test. Was evaluated, daily, the colony diameter and the mycelial mass obtained after five days of culture. The medium containing peanut husks extract provides the optimal development for the Lentinus sajor-caju strain, when an average of 68.3% of each petri dishes was colonized by mycelia. The medium containig castor bean seed shells were most favorable for Pleurotus ostreatoreoseus development, such colonizing 40,01% of each petri dishes. The wild strain of Pleurotus pulmonarius grew indifferently in the tested media and reaches the higher development when compared with Pleurotus ostreatoroseus strain. Lentinus sajor-caju demonstrate significantly more mycelial dry mass in the peanut shells medium when compared with the others strains. This result suggests that the mycelial growing of Lentinus sajor-caju and Pleurotus ostreatoroseus are influencied by the media. For the wild strain the mycelial growing its not significantly influencied by the tested media.

Obtenção de linhagens de Lentinula edodes resistentes a temperaturas elevadas e seleção de linhagens resistentes ao Trichoderma sp / Obtention of Lentinula edodes strains resistant to high temperatures and selection of strains resistant to Trichoderma sp

Lentinula edodes (Berk.) Pegler (shiitake) é o segundo cogumelo mais consumido no mundo e sua produção pode ser feita em toras de madeira ou em cultivo axênico utilizando resíduos agroindustriais como substrato. O cultivo do shiitake no Brasil tem sido gravemente prejudicado pela alta incidência de fungos contaminantes, como Trichoderma sp., e pela ausência de linhagens mais adaptadas às temperaturas tropicais. Duas linhagens resistentes ao Trichoderma sp. foram obtidas por meio de seleção quando crescidas em contato com o fungo contaminante e, por meio de fragmentação micelial, foram obtidas linhagens resistentes a temperaturas elevadas, capazes de crescer a 30º C em meio de cultura.

Lentinula edodes (Berk.) Pegler (shiitake) is the second most consumed mushroom in the world and its production can be made in natural logs or in axenic culture using agroindustrial residues as substrate. The cultivation of shiitake in Brazil has been seriously harmed by the high incidence of contaminant fungi such as Trichoderma sp. and by the absence of strains adapted to tropical temperatures. Two strains resistant to Trichoderma sp. were obtained by selection when grown in contact with the contaminant fungus and, by mycelial fragmentation, strains were obtained that can resist high temperatures and grow at 30º C in a culture medium.

Entomofauna associada ao cultivo de Pleurotus sajor-caju (Fr) Singer (Agaricales: Agaricaceae) no município do Capão do Leão, RS, Brasil / Insects associated with the Pleurotus sajor-caju (Fr) Singer (Agaricales: Agaricaceae) crop in the county of Capão do Leão, RS, Brazil

O objetivo deste trabalho foi conhecer a diversidade de artrópodes associada ao cultivo de Pleurotus sajor-caju em palha de arroz pasteurizada em ambiente semicontrolado, localizado no sul do Rio Grande do Sul. A captura dos espécimes foi realizada no período de abril a maio de 2007, durante a fase de frutificação da cultura, compreendendo 45 dias. Foram realizadas nove coletas com intervalos de 5 dias, utilizando armadilhas INTRAL AL 012 (12 volts). Capturou-se um total de 3.129 indivíduos divididos em cinco ordens com predominância de Diptera (2.874), Hymenoptera (152), Ortoptera (69), Colembola (59) e Coleoptera (38). A ordem Diptera foi a que apresentou o maior número de famílias, sendo Mycetophilidae (2.530), Sciaridae (264), Cecidomyiidae (10), Phoridae (46), Muscidae (4), Calliphoridae (1) e Culicidae (19).

The objective of this research was to know the diversity of arthropods associated to the Pleurotus sajor-caju crop in rice straw pasteurized in a semicontrolled environment, located in the south of the state of Rio Grande do Sul, Brazil. The capture of specimens was carried out in the period of April through May 2007, during the phase of fruit bodies of the crop, lasting 45 days. Nine collections with intervals of 5 days were carried out, using INTRAL AL 012 (12 volt) traps. A total of 3129 individuals divided into five orders with predominance of Diptera (2874), Hymenoptera (152), Ortoptera (69), Colembola (59) and Coleoptera (38) was captured. The Diptera order was the one that presented the biggest number of families, being Mycetophilidae (2530), Sciaridae (264), Cecidomyiidae (10), Phoridae (46), Muscidae (4), Calliphoridae (1) and Culicidae (19).

Cultivation of Agaricus blazei on Pleurotus spp. spent substrate

The aim of this work was the use of Pleurotus ostreatus and Pleurotus sajor-caju for the previous lignocellulolytic decomposition of banana tree leaf straw and the further use of the degraded straw as substrate for the culture of Agaricus blazei. For optimising the production of A. blazei in terms of yield (Y percent) and biological efficiency (BE percent), adjustments to the composition of the substrate were evaluated in a 2(5) experimental design. The following components were tested in relation to percent of substrate dry mass: urea (1 and 10 percent), rice bran (10 or 20 percent) or ammonium sulphate (0 or 10 percent), inoculum (10 or 20 percent) and the casing material (subsoil or burned rice husks). The best results (79.71 Y percent and 6.73 BE percent) were found when the substrate containing 10 percent of rice bran, without ammonium sulphate, inoculated with 20 percent and covered with subsoil was used.

O cultivo de fungos comestíveis e medicinais utilizando resíduos da agroindústria vem se apresentando como uma alternativa econômica para o pequeno produtor rural, favorecendo a agricultura familiar do nordeste catarinense. Este trabalho avaliou o fungo Pleurotus para a decomposição lignocelulolítica de palha de folhas de bananeira e a utilização da palha residual como substrato para o cultivo de Agaricus blazei. Ajustes na composição do substrato residual de Pleurotus, tais como o tipo e a concentração da fonte de nitrogênio, a porcentagem de inóculo e a camada de cobertura, foram avaliadas. O substrato residual que mais favoreceu a produção de A. blazei em Eficiência Biológica (6,73 por cento), Rendimento (79,71 por cento) e menor tempo para emissão do primeiro primórdio (27 dias) foi o substrato residual de P. ostreatus inoculado com 20 por cento de inóculo (ms), 10 por cento de farelo de arroz (ms), sem sulfato de amônio e utilizando terra de subsolo como camada de cobertura.

Isolamento e identificação de bactérias presentes nos solos de cobertura utilizados no cultivo do cogumelo Agaricus blazei Murril / Isolation and identification of bacteria present in the casing layer utilized to the cultivation of the mushroom Agaricus blazei Murril

Este trabalho foi desenvolvido com o objetivo de isolar e identificar bactérias presentes nos solos utilizados como camada de cobertura do cultivo de Agaricus blazei Murril. Foi avaliada a eficiência da desinfestação com vapor d'água e formol nos solos de cobertura. O tratamento com formol apresentou maior redução no número de bactérias na terra de barranco (Latossolo Vermelho distroférrico) e solo de pastagens (Latossolo Amarelo distrófico), enquanto que no solo hidromórfico, oriundo de várzea (Gleissolo Melânico), nenhum dos tratamentos foi eficiente para reduzir a comunidade microbiana. Foram identificadas bactérias do gênero Pseudomonas, Salmonella, Enterobacter e Shigella, dentre outras, denotando que os tratamentos de desinfestação não eliminam bactérias potencialmente benéficas, mas também não eliminam bactérias potencialmente patogênicas. Além disso, os tratamentos de desinfestação também não afetaram a produtividade do cogumelo A. blazei nas condições testadas.

The objective of this work was to quantify and to isolate the bacteria present in casing layer used for the cultivation of the Agaricus blazei Murril. It was evaluated the efficiency of the steam and formaldehyde disinfestation in the casing layer. The formaldehyde disinfestation showed a reduction of the bacteria population on the ravine soils (distroferric Red Latosol) and grazing land soil (distrofic Yellow Latosol), while in the hydromorphic soil coming from lowland (Melanic G]/leysol) none of the treatments used were efficient to reduce the bacteria population. Bacteria of the genera Pseudomonas, Salmonella, Enterobacter, Shigella, among others, were identified in all treatments analysed. It was observed that the disinfestation treatments did not eliminate neither potentially beneficial bacteria nor pathogenic bacteria. There was no significant effect of the disinfestation treatments on productivity of A. blazei mushroom. Moreover, the disinfestation treatments did not affect the productivity of the A. blazei in the studied conditions.