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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 KH 2 PO 4 , 0.5 g/L MgSO 4 , 0.001 g/L FeSO 4 , 0.01 g/L ZnSO 4 , and 0.01 g/L MnSO 4 . 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.

This study aimed to evaluate the effects of the solid and semisolid culture medium on the mycelial viability of A. subrufescens after 5-year cryopreservation at - 70 °C. Mycelia were grown in three types of whole or ground grains, with or without 5% glycerol addition in the substrate and/or in a cryotube. After 5 years of cryopreservation at - 70 °C, every treatment was thawed and recovered in malt extract culture medium with 15 (solid culture medium) or 5 g L -1 (semisolid culture medium) of agar. The semisolid recovery culture medium increased the mycelial viability recovery capacity of A. subrufescens cryopreserved for 5 years in grains with glycerol only in the cryotube, and specifically with medium-hard wheat grain without glycerol addition at all. Agar-based substrates such as malt extract agar, agar-ground grain, or the one with glycerol addition to the substrate were not effective to keep the mycelial viability, regardless of the recovery culture medium consistency. Hard and medium-hard endosperm wheat grains or hard endosperm rye grains with addition of glycerol as cryoprotectant only to the cryotube were effective to cryopreserve the fungus for 5 years without cryoprotectant addition in the substrate.

Gallesia integrifolia (Phytolaccaceae) is native to Brazil and has a strong alliaceous odor. The objective of this study was to identify the chemical composition of G. integrifolia fruit essential oil and evaluate fungicidal activity against the main food-borne diseases and food spoilage fungi. The essential oil was extracted by hydrodistillation and identified by GC-MS. From 35 identified compounds, 68% belonged to the organosulfur class. The major compounds were dimethyl trisulfide (15.49%), 2,8-dithianonane (52.63%) and lenthionine (14.69%). The utilized fungi were Aspergillus fumigatus, Aspergillus niger, Aspergillus ochraceus, Aspergillus versicolor, Penicillium funiculosum, Penicillium ochrochloron, Penicillium verrucosum var. cyclopium, and Trichoderma viride. Minimal fungicidal concentration for the essential oil varied from 0.02 to 0.18mg/mL and bifonazole and ketoconazole controls ranged from 0.20 to 3.50mg/mL. The lower concentration of the essential oil was able to control P. ochrochloron, A. fumigatus, A. versicolor, A. ochraceus and T. viride. This study shows a high fungicidal activity of G. integrifolia fruit essential oil and can support future applications by reducing the use of synthetic fungicides., (Copyright © 2018 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.)

Agaricus subrufescens is a basidiomycete which is studied because of its medicinal and gastronomic importance; however, less attention has been paid to its preservation. This study aimed to evaluate the effect of sucrose addition to substrate and cryotube on the viability of Agaricus subrufescens cryopreserved at -20°C and at -75°C for one and two years. Zero, 10% or 20% sucrose was added to potato dextrose agar or wheat grain. The mycelia were cryopreserved in the absence of cryoprotectant or with sucrose solutions at 15%, 30% or 45%. After one or two years at -75°C or at -20°C, mycelia were thawed and evaluated about viability, initial time of growth, colony diameter and genomic stability. Cryopreservation at -20°C is not effective to keep mycelial viability of this fungus. Cryopreservation at -75°C is effective when sucrose is used in substrates and/or cryotubes. Without sucrose, cryopreservation at -75°C is effective only when wheat grains are used. Physiological characteristic as mycelial colony diameter is negatively affected when potato dextrose agar is used and unaffected when wheat grain is used after two-year cryopreservation at -75°C. The fungus genome does not show alteration after two-year cryopreservation at -75°C., (Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.)

Basidiomycetes have several biotechnological and industrial applications such as enzyme production, bioremediation, pharmaceutical and functional food production. Due to climatic features, the preservation of several basidiomycetes is threatened, and to guarantee the preservation of this genetic resource, the development of long-term preservation techniques is necessary once there is no universal protocol for the cryopreservation of basidiomycetes. Cryopreservation is a technique in which microorganisms are submitted to ultralow temperatures. Therefore, this study aimed to collect information on the main conditions for long-term cryopreservation of basidiomycetes in the last 20 years. Scientific articles on cryopreservation of basidiomycetes published from 1997 to 2016, were researched, and only the studies on two intervals of cryopreservation were considered: from 1 to 2 years and for longer than 2 years. The analyzed conditions of basidiomycete cryopreservation were: most studied genera, cryopreservation temperature, substrate, cryoprotectant (and preservation substrate), cryopreservation period, thawing temperature and cultivation medium after thawing, physiological and genetic stability of basidiomycetes after thawing in cryopreservation. In this review, the viability of the main cryopreservation conditions of basidiomycetes studied in the last 20 years are presented and discussed., (Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.)

Pleurotus ostreatus is able to bioaccumulate several metals in its cell structures; however, there are no reports on its capacity to bioaccumulate iron. The objective of this study was to evaluate cultivation variables to increase iron bioaccumulation in P. ostreatus mycelium. A full factorial design and a central composite design were utilized to evaluate the effect of the following variables: nitrogen and carbon sources, pH and iron concentration in the solid culture medium to produce iron bioaccumulated in mycelial biomass. The maximum production of P. ostreatus mycelial biomass was obtained with yeast extract at 2.96 g of nitrogen L (-1) and glucose at 28.45 g L (-1) . The most important variable to bioaccumulation was the iron concentration in the cultivation medium. Iron concentration at 175 mg L (-1) or higher in the culture medium strongly inhibits the mycelial growth. The highest iron concentration in the mycelium was 3500 mg kg (-1) produced with iron addition of 300 mg L (-1) . The highest iron bioaccumulation in the mycelium was obtained in culture medium with 150 mg L (-1) of iron. Iron bioaccumulation in P. ostreatus mycelium is a potential alternative to produce non-animal food sources of iron.

The essential oil of Lippia alba is reported as an antifungal against human pathogenic microorganisms but few articles report its use as an alternative to synthetic fungicides on green mould control. The objective of this study was to determine chemical characteristics of L. alba essential oil and its antifungal activity against green molds as an alternative to synthetic fungicides. Essential oil was extracted by Clevenger hydrodistillation, characterized by GC-MS analysis, and the structure of the main compounds confirmed by (1)H and (13)C-NMR spectroscopy. Microdilution assays evaluated the essential oil minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC). Commercial fungicides Ketoconazole and Bifonazole were used as control. Essential oil yield is of 0.15% and the major components are neral (33.32%) and geranial (50.94%). The L. alba essential oil has MIC of 0.300-1.250 mg/mL and MFC of 0.600-1.250 mg/mL. Ketoconazole and Bifonazole show MIC ranging from 0.025-0.500 to 0.100-0.200 mg/mL, and MFC ranging from 0.250-0.100 to 0.200-0.250 mg/mL, respectively. L. alba essential oil is classified as citral type and the results indicate that it is a potential alternative to synthetic fungicides.

This study aimed to verify the biological efficiency and production flushes of Agaricus blazei strains on different casing layers during 90 cultivation days. Four casing layers were used: mixture of subsoil and charcoal (VCS), lime schist (LSC), São Paulo peat (SPP) and Santa Catarina peat (SCP); and two genetically distant A. blazei strains. The fungus was grown in composted substratum and, after total colonization, a pasteurized casing layer was added over the substratum, and fructification was induced. Mushrooms were picked up daily when the basidiocarp veil was stretched, but before the lamella were exposed. The biological efficiency (BE) was determined by the fresh basidiocarp mass divided by the substratum dry mass, expressed in percentage. The production flushes were also determined over time production. The BE and production flushes during 90 days were affected by the strains as well as by the casing layers. The ABL26 and LSC produced the best BE of 60.4%. Although VCS is the most used casing layer in Brazil, it is inferior to other casing layers, for all strains, throughout cultivation time. The strain, not the casing layer, is responsible for eventual variations of the average mushroom mass. In average, circa 50% of the mushroom production occurs around the first month, 30% in the second month, and 20% in third month. The casing layer water management depends on the casing layer type and the strain. Production flush responds better to water reposition, mainly with ABL26, and better porosity to LSC and SCP casing layers.

The fungus Agaricus brasiliensis is a Basidiomycete studied because of its immunomodulation and/or antitumor substances. The objective of this study was to verify the Agaricus brasiliensis antineoplasic activity in vivo on different basidiocarp maturation phases on Sarcoma 180 cells implanted in mice. Sarcoma cells were implanted in mice and after seven days mice were divided in three groups. The first group was treated with saline solution, the second group was treated with closed basidiocarp extract solution and the third group was treated with opened basidiocarp extract solution. After 30 days of being daily orally treated with these three solutions all animals suffered euthanasia, and the splenic index, tumor mass and volume were determined. No significant differences of the tumor growth inhibition in function of the different basidiocarp maturation phases for the Agaricus brasiliensis strain were observed. The in vivo basidiocarp antineoplasic average activity was 89.22%.