Photoregulation of the biosynthetic activity of the edible medicinal mushroom Lentinula edodes in vitro.

The findings of the study demonstrate the impact of low-intensity laser and quasi-monochromatic light on the biosynthetic activity of the edible medicinal fungus L. edodes during submerged cultivation. An artificial lighting installation based on matrices of light-emitting diodes (LED) emitting light at 470 nm (blue), 530 nm (green), 650 nm (red), and argon gas laser (488 nm) was used. Irradiation with blue and red LED and laser led to a shortening of the lag phase by 2 days and an increase in the mycelial mass. Irradiation with laser light resulted in the highest mycelial mass yield (14.1 g/L) on the 8th day of cultivation. Irradiation in all used wavelength ranges caused an increase in the synthesis of both extracellular and intracellular polysaccharides. Laser light at 488 nm and LED at 470 nm proved to be the most effective. Irradiation with red, green, and blue laser light caused an increase in the total amount of fatty acids in the mycelial mass compared to the control. A significant distinction in qualitative composition was observed: short-chain acids C6‒C12 compounds were produced under red light irradiation, whereas long-chain C20‒C24 were formed under green light irradiation. The most significant changes in the aromatic profile of the mycelial mass and culture liquid were recorded upon irradiation with green light. The content of aromatic components increased 24.6 times in the mycelial mass and 38.5 times in the culture liquid. The results suggest the possibility of using low-intensity quasi-monochromatic light for targeted regulation of L. edodes biosynthetic activity.

Characterization, In Vitro Biological Activity and In Vivo Cardioprotective Properties of Trametes versicolor (L.:Fr.) Quél. Heteropolysaccharides in a Rat Model of Metabolic Syndrome.

The present study aimed to examine the biological activity and cardioprotective potential of Trametes versicolor heteropolysaccharides (TVH) in a rat model of metabolic syndrome (MetS). This study included 40 Wistar rats divided into 5 groups: CTRL-healthy non-treated rats; MetS-non-treated rats; and H-TV, M-TV and L-TV-rats with MetS treated with either 300, 200 or 100 mg/kg TVH per os for 4 weeks. After finishing the treatment, we conducted an oral glucose tolerance test (OGTT), hemodynamic measurements and the animals were sacrificed, hearts isolated and subjected to the Langendorff technique. Blood samples were used for the determination of oxidative stress parameters, lipid status and insulin levels. We showed that α-amylase inhibition was not the mode of TVH antidiabetic action, while TVH showed a moderate inhibition of pathogenic microorganisms' growth (MIC 8.00 mg·mL-1; MBC/MFC 16.00 mg·mL-1). H-TV and M-TV significantly reduced the level of prooxidants (O2-, H2O2, TBARS; p < 0.05), increased antioxidants activity (SOD, CAT, GSH; p < 0.05), reduced blood pressure (p < 0.05), improved glucose homeostasis in the OGTT test (p < 0.05), and ejection fraction (p < 0.05) and cardiac contractility (p < 0.05) compared to MetS (p < 0.05). Moreover, TVH treatment normalized the lipid status and decreased insulin levels compared to MetS rats (p < 0.05). The obtained results demonstrated that the TVH may be considered a useful agent for cardioprotection in MetS conditions.

Cultivation Technologies for Production of Medicinal Mushroom Biomass: Review.

Since most medicinal mushrooms are rare in nature, artificial cultivation and farming of their fruiting bodies as well as biotechnological cultivation of their fungal biomass in bioreactors on solid substrates and in liquid media has been established. This review compares the benefits and weaknesses of all three capabilities. Cultivation technologies are presented, including traditional cultivation via farming of fruiting bodies on wood logs and on sawdust-based substrates and modern biotechnological cultivation of mycelia in bioreactors by submerged and solid-state cultivation technologies. Our findings indicate that farming is cost-effective but large-scale production is time-consuming. In addition, solid-state cultivation is a comprehensive well-controlled technology that is close to the natural growth process and is suitable for veterinary use and use in food supplements. Finally, submerged liquid-state cultivation of mushroom mycelia is fast and well controlled.

Culture Degeneration Reduces Sex-Related Gene Expression, Alters Metabolite Production and Reduces Insect Pathogenic Response in Cordyceps militaris.

Cordyceps militaris is an entomopathogenic ascomycete, known primarily for infecting lepidopteran larval (caterpillars) and pupal hosts. Cordycepin, a secondary metabolite produced by this fungus has anti-inflammatory properties and other pharmacological activities. However, little is known about the biological role of this adenosine derivate and its stabilising compound pentostatin in the context of insect infection the life cycle of C. militaris. During repeated subcultivation under laboratory conditions a degeneration of C. militaris marked by decreasing levels of cordycepin production can occur. Here, using degenerated and parental control strains of an isolate of C. militaris, we found that lower cordycepin production coincides with the decline in the production of various other metabolites as well as the reduced expression of genes related to sexual development. Additionally, infection of Galleria mellonella (greater wax moth) caterpillars indicated that cordycepin inhibits the immune response in host haemocytes. Accordingly, the pathogenic response to the degenerated strain was reduced. These data indicate that there are simultaneous changes in sexual reproduction, secondary metabolite production, insect immunity and infection by C. militaris. This study may have implications for biological control of insect crop pests by fungi.

Novel Medicinal Mushroom Blend as a Promising Supplement in Integrative Oncology: A Multi-Tiered Study using 4T1 Triple-Negative Mouse Breast Cancer Model.

Although medicinal mushroom extracts have been proposed as promising anti-cancer agents, their precise impacts on metastatic breast cancer are still to be clarified. For this purpose, the present study exploited the effect of a novel medicinal mushroom blend, namely Micotherapy U-care, in a 4T1 triple-negative mouse breast cancer model. Mice were orally administered with Micotherapy U-care, consisting of a mixture of Agaricus blazei, Ophiocordyceps sinensis, Ganoderma lucidum, Grifola frondosa, and Lentinula edodes. The syngeneic tumor-bearing mice were generated by injecting 4T1 cells in both supplemented and non-supplemented mice. After sacrifice 25 days later, specific endpoints and pathological outcomes of the murine pulmonary tissue were evaluated. (i) Histopathological and ultrastructural analysis and (ii) immunohistochemical assessment of TGF-ß1, IL-6 and NOS2, COX2, SOD1 as markers of inflammation and oxidative stress were performed. The QoL was comparatively evaluated. Micotherapy U-care supplementation, starting before 4T1 injection and lasting until the end of the experiment, dramatically reduced the pulmonary metastases density, also triggering a decrease of fibrotic response, and reducing IL-6, NOS, and COX2 expression. SOD1 and TGF-ß1 results were also discussed. These findings support the valuable potential of Micotherapy U-care as adjuvant therapy in the critical management of triple-negative breast cancer.

Comparison of lovastatin, citrinin and pigment production of different Monascus purpureus strains grown on rice and millet.

Six Monascus purpureus (red mould) strains were cultivated on brown rice and millet as substrates. They underwent strain selection for high lovastatin and pigment production, and low citrinin mycotoxin production, with particular reference to potential for millet as substrate. For most of these strains, substrate dry matter loss was 54-60% on rice and 46-48% on millet, although the 'MOPU GS1' strain showed 18% and 17% dry matter loss, respectively. 'MOPU GS1' was also the only strain with detectable levels of lovastatin (1.3 and 1.6 mg lovastatin/g substrate dry weight [dw], respectively) and citrinin under the limit of detection. In the other strains, citrinin varied from 0.3 to 18.2 mg/g substrate dw. Among the six strains, 'EBY3' provided high pigment production when grown on rice, although it produced 1.1 mg citrinin/g substrate dw. Millet showed good potential as an alternative substrate to rice, due to higher lovastatin and lower citrinin production; however, rice was the better substrate for production of M. purpureus pigments.

Dietary Supplementation of Hericium erinaceus Increases Mossy Fiber-CA3 Hippocampal Neurotransmission and Recognition Memory in Wild-Type Mice.

Hericium erinaceus (Bull.) Pers. is a medicinal mushroom capable of inducing a large number of modulatory effects on human physiology ranging from the strengthening of the immune system to the improvement of cognitive functions. In mice, dietary supplementation with H. erinaceus prevents the impairment of spatial short-term and visual recognition memory in an Alzheimer model. Intriguingly other neurobiological effects have recently been reported like the effect on neurite outgrowth and differentiation in PC12 cells. Until now no investigations have been conducted to assess the impact of this dietary supplementation on brain function in healthy subjects. Therefore, we have faced the problem by considering the effect on cognitive skills and on hippocampal neurotransmission in wild-type mice. In wild-type mice the oral supplementation with H. erinaceus induces, in behaviour test, a significant improvement in the recognition memory and, in hippocampal slices, an increase in spontaneous and evoked excitatory synaptic current in mossy fiber-CA3 synapse. In conclusion, we have produced a series of findings in support of the concept that H. erinaceus induces a boost effect onto neuronal functions also in nonpathological conditions.

Effect of Solid State Fermentation Medium Optimization on Pleurotus ostreatus Laccase Production.

The objective of this work was to increase laccase production by Pleurotus ostreatus PLAB through culture medium optimization using solid state culture conditions. Increased laccase activity was obtained through design of experiments (DOE) using the Taguchi orthogonal array (OA). Seven factors, viz. lignocellulose, glucose, yeast extract, peptone, KH(2)PO(4), MgSO(4) 7H(2)O and MnSO(4) H(2)O at three levels and pH at two levels. OA layout of L18 (2(1) x 3(7)) was selected for the proposed experimental design using Minitab 17 software. Data analysis showed that lignocellulose (20 %) and glucose (10 g L1) had positive effect, whereas KH(2)PO(4), MgSO(4)∙7H(2)O and MnSO(4)∙H(2)O did not have significant effect on laccase production. Taguchi OA analysis showed that pH 6, lignocellulose 20 %, glucose 10 g L(-1), yeast extract 6 g L(-1), peptone 15 g L(-1), KH(2)PO(4) 3 g L1, MgSO(4)∙7H(2)O 0.5 g L(-1) and MnSO(4)∙H(2)O 0.1 g L-1 were the optimal conditions to maximize laccase production. The model predicted a 30.37 U g(-1) dry wt., which agreed with the experimentally obtained laccase activity 29.15 U g(-1) dry wt. at optimal conditions.

Immunomodulating activities of cultivated maitake medicinal mushroom Grifola frondosa (Dicks.: Fr.) S.F. Gray (higher Basidiomycetes) on peripheral blood mononuclear cells.

Grifola frondosa is a culinary-medicinal mushroom that contains several physiologically active compounds, of which polysaccharides, specifically ß-glucans, are known to possess immunomodulating properties. Its extracts are studied for application as adjuncts for chemotherapy, and experiments in animal models support the use of this mushroom for cancer treatment. The effect of extracts obtained from mushrooms cultivated on different substrates and their capacity of inducing the secretion of cytokines from human peripheral blood mononuclear cells were studied. The activity of extracts at concentrations 12.5, 100, and 200 µg/mL on induction of TNF-α, IFN-γ, and IL-12 was screened. Two extracts from substrates fortified with olive oil press cakes showed appreciable activity and induced the secretion of TNF-α, IL-12, and INF-γ. The extracts differed from the others in the amount of sugar, protein, and ß-glucans, which can explain their higher activity. Present results show that different substrates and different source materials can reasonably modify the bioactivity of cultivated G. frondosa.

Influence of olive oil press cakes on Shiitake culinary-medicinal mushroom, lentinus edodes (Berk.) singer (higher basidiomycetes) fruiting bodies production and effect of their crude polysaccharides on CCRF-CEM cell proliferation.

Lentinus edodes (Berk.) Singer fruiting bodies were cultivated on substrates composed of beech sawdust, wheat bran, and calcium sulfate hemihydrate (gypsum), containing different proportions of olive oil press cakes (OOPC). We determined the influence of OOPC on fruiting bodies production and proliferation of CCRF-CEM leukemia cells. A negative influence of OOPC on mycelia growth and maturation was noticed. When growth medium contained 80% OOPC, fruiting bodies ceased forming. To investigate the cytotoxicity on CCRF-CEM cells in vitro, cells were treated with crude polysaccharides extracted from L. edodes fruiting bodies. Also in this case a negative correlation between OOPC content and cytotoxicity was found.

Cultivation and bioactivity assessment of Grifola frondosa fruiting bodies on olive oil press cakes substrates.

Grifola frondosa, a medicinal and edible mushroom, was cultivated on substrates composed of olive oil press cakes with different supplements. Crude extracts were prepared from mature fruiting bodies and tested for their capacity to stimulate splenocyte proliferation. Two wild-growing fruiting bodies were extracted for comparison. Olive oil press cakes reduced the mushroom yield, and the best biological efficiency was obtained on substrates supplemented with wheat bran and without olive oil press cakes. All extracts were capable of inducing splenocyte proliferation and were half as effective as the positive control (6.0 microg/mL phytohaemagglutinin). No correlation between substrate composition and bioactivity could be established. Extracts from wild-growing G. frondosa were superior to cultivated ones in respect to biological activity.

The use of spent brewery grains for Pleurotus ostreatus cultivation and enzyme production.

In the brewing industry, spent brewery grains (SBGs) are byproducts with a low economic value. The potential use of this leftover as a substrate ingredient for Pleurotus ostreatus fruiting body cultivation and enzyme production was evaluated. The best substrate mixture for P. ostreatus mycelium growth comprised 30% wheat bran (WB), 68% beech sawdust (BS) and 2% CaCO3. On the substrates containing SBG, the fastest mycelium growth was observed on the substrate composed of 10% SBG, 20% WB, 68% BS and 2% CaCO3. The highest biological efficiency (51%) of fruiting bodies was determined on the mixtures containing 20% WB, 10% SBG and 2% CaCO3. The SBGs with the addition of WB were also shown to be suitable as a substrate for enzyme production. However, the supplementation levels designate which enzymes are produced and in what amounts.