Pachira aquatica: biological activity and chemical composition of leaves, flowers, and seeds.

The chemical composition of Pachira aquatica crude extracts flowers, leaves, and seeds was obtained by UHPLC-ESI/qTOF and GC/MS. The antiproliferative activity was evaluated against the human tumour cell lines AGS (gastric), CaCo-2 (colorectal), MCF-7 (breast), and NCI-H460 (lung). The anti-inflammatory and cellular antioxidant activities were also studied. Flavonoids, phenolic acids, coumarins, and saturated fatty acids were identified in the samples. The concentration of extracts responsible for inhibiting 50% of nitric oxide production ranged from (149 to > 400 µg mL-1). Antiproliferative activity against the tumour cell lines was: AGS (GI50 175 to > 400 µg mL-1), Caco-2 (GI50 215 to > 400 µg mL-1), MCF7 (GI50 232 to > 400 µg mL-1) and NCI-H460 (GI50 208 to > 400 µg mL-1). Cellular antioxidant activity remained between 73% to > 2000%. The selectivity index (SI) ranged from 1.00 to 2.78, indicating low antiproliferative activity.

Cellular Antioxidant, Anti-Inflammatory, and Antiproliferative Activities from the Flowers, Leaves and Fruits of Gallesia integrifolia Spreng Harms.

Gallesia integrifolia, a notable species in the Atlantic Forest, has been traditionally employed in folk medicine for treating rheumatism, asthma, and worms. This study investigated the cellular antioxidant, antiproliferative, and anti-inflammatory activities of the essential oils (EOs) and crude extracts (CEs) from G. integrifolia flowers, fruits, and leaves. The chemical identification of EOs was performed by GC-MS and CEs by UHPLC-MS. Cellular antioxidant and anti-inflammatory activities were assessed through mouse macrophage cell culture. In addition, the antiproliferative potential was evaluated in gastric, colorectal, breast, and lung tumor cell lines and non-tumor VERO cells. EOs predominantly contained organosulfur compounds in flowers (96.29%), fruits (94.94%), and leaves (90.72%). We found the main compound is 2,2'-Disulfanediyldiethanethiol in the EOs of flowers (47.00%), leaves (41.82%), and fruits (44.39%). Phenolic compounds were identified in CEs. The EOs and CEs demonstrated potential against the tumor cell lines tested (GI50 between 51 and 230 µg/mL). The selectivity index values were greater than 1.0 (1.01 to 3.37), suggesting a relative safety profile. Moreover, the anti-inflammatory activity IC50 ranged from 36.00 to 268 µg/mL, and the cellular oxidation inhibition ranged from 69% to 82%. The results suggest that oils and extracts derived from G. integrifolia have potential for use in various industrial sectors.

Biosorption of methylene blue by residue from Lentinus crinitus mushroom cultivation.

Conventional textile effluent treatments cannot remove methylene blue, a mutagenic azo dye, and an endocrine disruptor, that remains in the drinking water after conventional water treatment. However, the spent substrate from Lentinus crinitus mushroom cultivation, a waste, could be an attractive alternative to remove persistent azo dyes in water. The objective of this study was to assess the methylene blue biosorption by spent substrate from L. crinitus mushroom cultivation. The spent substrate obtained after mushroom cultivation had been characterized by the point of zero charge, functional groups, thermogravimetric analysis, Fourier transform infrared spectroscopy, and scanning electron microscopy. Moreover, the spent substrate biosorption capacity was determined in function of pH, time, and temperature. The spent substrate had a point of zero charge value of 4.3 and biosorbed 99% of methylene blue in pH from 3 to 9, with the highest biosorption in the kinetic assay of 15.92 mg g- 1, and in the isothermal assay of 120.31 mg g- 1. Biosorption reached equilibrium at 40 min after mixing and best fitted the pseudo-second-order model. Freundlich model best fitted the isothermal parameters and each 100 g spent substrate biosorbed 12 g dye in an aqueous solution. The spent substrate of L. crinitus cultivation is an effective biosorbent of methylene blue and an alternative to removing this dye from water, adding value to the mushroom production chain, and supporting the circular economy.

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.

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.

Long-term cryopreservation of Lentinus crinitus strains by wheat grain technique.

Lentinus crinitus (Basidiomycota: Polyporales) is a saprophytic fungus with biotechnological importance described more than 20 years ago. However, there are few studies on the long-term preservation of this basidiomycete. Cryopreservation is a long-term storage technique that reduces the metabolic activity of microorganisms, but its success depends on the adjustment of the freezing process, the cryoprotectants, and the protective substrates for each species. This study aimed to assess the mycelial viability and genetic stability of L. crinitus strains cryopreserved at -86 °C for two years by the wheat grain technique using different cryoprotectants and freezing methods. Three strains of L. crinitus (U9-1, U13-5, and U15-12) were subjected to different concentrations and types of cryoprotectants (dimethyl sulfoxide, glycerol, glucose, and sucrose), freezing methods such as immediate freezing from 25 to -86 °C and progressing freezing from 25 to -86 °C in a freezing container with isopropyl alcohol to control the rate of cell freezing at -1 °C min-1, protective substrate (wheat grain and 2% malt extract agar), and cryopreservation period (1, 6, 12, and 24 months). After thawing, samples were evaluated for mycelial viability, time to mycelial recovery, mycelial stability, and genetic stability of the fungus. All techniques achieved effective cryopreservation at -86 °C, mainly with the wheat grain technique. All cryoprotectants (3.5% glycerol, 1.5% dimethyl sulfoxide, 25% sucrose, and 5% glucose), freezing methods (immediate and gradual), and protective substrate (wheat grain and malt extract agar) were effective for cryopreservation of the three L.crinitus strains in an ultra-low temperature freezer for two years. Mycelial viability, mycelial stability, and genetic stability of the fungus were not affected after two-year cryopreservation, evidencing the robustness of the long-term cryopreservation technique and the fungus.

Endophytic fungi of Brunfelsia uniflora: isolation, cryopreservation, and determination of enzymatic and antioxidant activity.

Brunfelsia uniflora (Pohl.) D. Don (Solanaceae), commonly known as manacá-de-cheiro, is widely distributed in Brazil and used by local indigenous peoples as an antirheumatic, antisyphilitic, depurative, emetic, vermifuge, and purgative agent. Several studies have examined the biological activities and phytochemical profile of Brunfelsia; however, few have focused on the diversity of endophytic microorganisms that colonize members of the genus. This study aimed to isolate and cryopreserve endophytic fungi from B. uniflora and determine their cellulase, laccase, and antioxidant activities. Endophytic fungi were isolated from B. uniflora stems, cultured on wheat grains, immersed in a 150 g L-1 aqueous sucrose solution, and cryopreserved at - 80 °C for 1 and 6 months. Cellulase activity was determined by a qualitative test using carboxymethylcellulose medium and laccase activity by a quantitative test based on the oxidation of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate). Prior to antioxidant activity assays, fungi were grown in malt extract broth for production of mycelial biomass. A methanolic extract was prepared for evaluation of DPPH· scavenging activity, FRAP activity, and total phenolic content. A total of 46 endophytic fungal isolates were obtained from B. uniflora stems and classified into 24 groups according to morphological similarities. B. uniflora was shown to harbor different genera of ascomycete fungi as endophytic organisms. Mycelial viability was observed after 1 and 6 months of cryopreservation at - 80 °C. Fungi exhibited cellulase and laccase activities. Isolate CE23 had the highest laccase activity after 7 days of cultivation. Twelve isolates were found to have low total phenolic contents and DPPH· and FRAP activities.

Basidiocarp structures of Lentinus crinitus: an antimicrobial source against foodborne pathogens and food spoilage microorganisms.

Lentinus crinitus basidiocarps are an alternative to antimicrobials, but the stipe (24% basidiocarp) is discarded even with potential antimicrobial activity. This study evaluated the antimicrobial activity of L. crinitus basidiocarp pileus and stipe extracts against foodborne pathogens and food spoilage microorganisms. Basidiocarps of L. crinitus were grown in sugarcane bagasse and rice husks and the pileus and stipe methanolic extract was analyzed by broth microdilution method for antimicrobial activity against eight bacteria and eight fungi. The minimum bactericidal concentration values for pileus and stipe ranged from 0.40 to 0.50 mg mL- 1, for streptomycin from 0.10 to 0.50 mg mL- 1, and for ampicillin from 0.40 to 1.20 mg mL- 1. The minimum fungicidal concentration values for pileus and stipe ranged from 0.06 to 0.60 mg mL- 1, for bifonazole from 0.20 to 0.25 mg mL- 1, and for ketoconazole from 0.30 to 3.50 mg mL- 1. Extracts had bacteriostatic, bactericidal, fungistatic and fungicidal activity against all microorganisms, but with greater efficiency and specificity for some microorganisms. Both pileus and stipe are promising and sustainable alternatives for use in food, agricultural, and pharmaceutical industries.

Iron-enriched mycelia of edible and medicinal basidiomycetes.

Iron bioaccumulation in basidiomycetes is an alternative to recover ferrous sulphate from titanium dioxide pigment production and to produce an iron-enriched mycelial biomass. This study aimed to evaluate iron bioaccumulation capacity in vegetative mycelium of edible and medicinal fungi grown in malt extract liquid medium with different ferrous sulphate contents. Five basidiomycetes were grown in malt extract liquid medium with different iron contents from 0.116 to 100 mg L-1 iron. The iron content of dried mycelial biomass bioaccumulated with iron was determined by flame atomic absorption spectrophotometry. All fungi grew on the iron culture media and the mycelial biomass growth ranged from 3.24 ± 0.65a mg mL-1 to 12.46 ± 0.29 mg mL-1. Iron addition to culture media increased the iron content in the mycelial biomass from 4000-13,000-fold compared with control. Pleurotus ostreatus (2181 ± 218 mg kg-1) presented the greatest iron content in the mycelial biomass, followed by Schizophyllum commune (1769 ± 131 mg kg-1), Agaricus subrufescens (1272 ± 8.84 mg kg-1), and Ganoderma lucidum (840 ± 75 mg kg-1). P. ostreatus, followed by S. commune, and G. lucidum at 90 and 100 mg L-1 iron in the culture medium are the best choices to produce iron-enriched mycelial biomass. This extensive study of several edible and medicinal basidiomycetes grown in different iron contents was effective in recovering ferrous sulphate byproduct and transferring it to mycelium to produce a new nutraceutical food of iron-enriched mycelial biomass.

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.

Iron biofortification and availability in the mycelial biomass of edible and medicinal basidiomycetes cultivated in sugarcane molasses.

Basidiomycetes can bioaccumulate high iron contents, but there are few studies on iron availability from the mycelial biomass in order to support their use as an iron-enriched fungal food. This study aimed to evaluate the in vitro iron bioaccumulation and availability in the mycelial biomass of edible and medicinal basidiomycetes grown in two distinct culture media. Lentinus crinitus, Ganoderma lucidum, Schizophyllum commune, Pleurotus ostreatus, Pleurotus eryngii, Lentinula edodes, and Agaricus subrufescens were grown in liquid culture medium of malt extract or sugarcane molasses to obtain iron-bioaccumulated mycelial biomass. P. ostreatus was the fungus that most bioaccumulated iron, followed by S. commune, and P. eryngii; they also had the highest mycelial biomass growth and iron transfer from the culture medium to the mycelial biomass. Mycelial iron availability is species-specific, regardless of the culture medium and the iron bioaccumulation capacity of the fungus in the mycelial biomass. Mycelial biomass of S. commune, followed by G. lucidum, P. ostreatus, and P. eryngii, associated with molasses culture medium, are the best choice for the production of iron-enriched mycelial biomass.

Five-year cryopreservation at -80 °C of edible and medicinal basidiomycetes by wheat grain technique.

This research has focused on basidiomycete cryopreservation at -80 °C and developed a cryopreservation method based on the use of hard or medium-hard endosperm wheat grains as a mycelial carrier for cryopreservation. The aim of this study was to evaluate the mycelial viability of edible and medicinal basidiomycetes, using 13 strains of Agaricus spp. and eight strains of non-Agaricus spp., cryopreserved at -80 °C on hard endosperm wheat grain, with or without cryoprotectant agent (4% glucose), for two and five years. Two groups of basidiomycetes, Agaricus genus and other non-Agaricus genera, were cryopreserved at -80 °C by wheat grain technique for two and five years. The cryopreservation technique with hard endosperm wheat grain without cryoprotectant (preservation substrate), settled previously for A. subrufescens is efficient to cryopreserve other basidiomycetes such as Lentinus crinitus, Pleurotus ostreatus, Pleurotus eryngii, Schizophyllum commune, and Lentinula edodes, besides A. subrufescens strains.

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.

Semisolid culture medium improves mycelial recovery of Agaricus subrufescens cryopreserved in cereal grains.

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.

Bioaccumulation of Lithium (Li2CO3) in Mycelia of the Culinary-Medicinal Oyster Mushroom, Pleurotus ostreatus (Agaricomycetes).

Pleurotus ostreatus is a white-rot mushroom that bioaccumulates metals in basidiocarps and vegetative mycelia. This fungus has been used in soil and water bioremediation of several heavy metals; however, there are few studies of lithium mycelial bioaccumulation for pharmacological use. The aim of this study was to evaluate lithium bioaccumulation in P. ostreatus mycelia grown in a liquid malt extract cultivation medium with Li2CO3 or LiCl. Each lithium source was added to the medium to obtain a concentration of 0, 5, 10, 15, 20, 25, 30, 40, 50, 100, or 200 mg · L-1 lithium. The highest bioaccumulation of lithium in mycelia was 1575.29 µg · g-1 upon treatment with 40 mg · L-1 Li2CO3. P. ostreatus mycelia produce biomass and bioaccumulate both lithium sources, but more lithium bioaccumulates when in the form of Li2CO3. This study provides a prospective for the development of biotechnological products with high aggregate values and alternative ways to deliver lithium and eventually other salts for pharmacological use.

Mycelial antineoplastic activity of Agaricus blazei.

Basidiocarp of Agaricus blazei (=Agaricus brasiliensis; =Agaricus subrufescens) is used as teas or capsules due to its antineoplastic effect but there are few reports of using mycelium for this purpose. The objective of this study was to evaluate the antineoplastic activity on sarcoma 180 cells implanted in mice of two forms of preparation of the mycelium from two A. blazei strains grown in culture medium with different concentrations of isolated soy protein. Mycelia were grown in Pontecorvo medium with different concentrations of isolated soybean protein (ISP). Mycelial hot water extract, moistened mycelial powder, hot water extract of green tea, Ifosfamida(®) (ifosfamide drug), and saline solution were administered daily by gavage in mice with sarcoma 180 cells to evaluate antineoplastic activity. It was concluded that antineoplastic activity was the same for both strains, except when used as moistened mycelial powder, which rules out the use of mycelial powder in capsules. Mycelial hot water extract had high antineoplastic activity with lower metabolic demand on the spleen and maintenance of normal blood parameters. Mycelial growth in different ISP concentrations had the same antineoplastic activity. Also the vegetative mycelium was as effective as the basidiocarp for sarcoma 180 tumor inhibition. Green tea was as effective as mycelial hot water extract.

Cryopreservation at -20 and -70 °C of Pleurotus ostreatus on Grains.

Alternative substrates for cryopreservation at -20 °C have been little explored for basidiomycetes and could bring new possibilities of lower cost cryopreservation. Nevertheless, freezing temperatures between -15 and -60 °C are very challenging because they frequently result in cryoinjuries. The objective of this study was to evaluate substrates associated to cryoprotective agents for Pleurotus ostreatus cryopreservation at -20 or -70 °C in order to develop alternative techniques for basidiomycete cryopreservation. P. ostreatus was grown on potato dextrose agar or whole grains of oat, wheat, rice or millet and transferred to cryovials with cryoprotective solution with 1 % dimethyl sulfoxide, 5 % glycerol, 10 % saccharose, 4 % glucose, 6 % polyethylene glycol-6000 or 5 % malt extract. The mycelium in the cryovials were cryopreserved at -20 or -70 °C and recovered for evaluation of the mycelial growth viability after 1 and 3 years. Both substrates and cryoprotectants affect the viability of the mycelial growth cryopreserved at -20 or -70 °C; wheat grains combined with cryoprotectants such as saccharose or glucose are effective for keeping mycelium viable after cryopreservation at -20 °C for 1 or 3 years; for cryopreservation at -70 °C after 1 or 3 years, any substrate combined with any cryoprotectant is effective for preserving the mycelium viable, except for millet grains with polyethylene glycol after 3 years; semi-permeable cryoprotective agents such as saccharose and glucose are the most effective for cryopreservation at -20 or -70 °C for at least 3 years.

Effects of Agaricus brasiliensis mushroom in Walker-256 tumor-bearing rats.

Agaricus brasiliensis is a mushroom native to São Paulo State, Brazil, that is studied for its medicinal proprieties. This work aimed to investigate the antitumoral activity of A. brasiliensis extracts and pure powdered basidiocarp preparation using Walker-256 (W256) tumor-bearing rats, a model for cancer-related cachexia studies. The rats were treated for 14 days by gavage (136 mg/kg) and at the end of the experiment tumors were collected to calculate mass and volume. Blood was collected for determination of plasma glucose, albumin, alanine aminotransferase (ALT), and aspartate aminotransferase (AST). Hepatic and tumor enzymes indicating oxidative stress were also evaluated. The results showed that all 4 treatments (pure powdered basidiocarp and aqueous, acid, and alkaline extracts) significantly reduced tumor size and promoted gain in body weight. Plasmatic analysis showed a reduction in AST level and increased glycemia in the treated rats. Pure basidiocarp preparations improved the liver catalase and superoxide dismutase activity, but did not change the glutathione S-transferase activity. The data collected from the W256 tumor-bearing rats revealed the beneficial effects of A. brasiliensis in tumor treatment, mainly related to cachexia. The benefits can be partly related to antioxidant activity and to reduction of weight loss and tumor growth.