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Abstract
The effect of lignocellulose and lignin on growth of mycelium of mushroom Lentinula edodes and laccase activity in cultivation medium was studied. It was shown that cultivation of L. edodes mycelium in liquid nutrient medium with addition of 0.25-0.5% of kraft lignin increased mycelium biomass yield approximately two times compared with reference conditions without addition of lignin. Similar results were obtained in experiments in which 0.5% lignocellulose that remained after obtaining furfural, and 0.5% lignin that remained after obtaining furfural and glucose from wheat straw, were added to the nutrient medium. This effect was greater in the conditions of cultivation with good aeration, compared with static culture. Laccase activity in medium increased after addition of wheat straw lignocellulose or lignin only in the case of mycelium cultivation with aeration. In the case of mushroom cultivation on solid nutrient medium, addition of wheat straw lignocellulose and lignin promoted growth of mycelium only during the first 7 days of cultivation.

The Cordyceps genus is well known for its medicinal properties. A number of bioactive constituents from Cordyceps species have been reported such as antibacterial, antifungal, immunopotentiating or antitumor agents [1]. The anti-inflammatory and anti-angiogenic properties of Cordyceps militaris (L.) Link have been reported as also the antioxidant activity of its cultured mycelium [2]. In this work, the chemical characterization of C. militaris was performed, includingbioactive compounds (free sugars, unsaturated fatty acids, tocopherols, organic acidsand phenolic compounds). The antioxidant potential of its methanolic extract was evaluated (reducing power, scavenging activity and lipid peroxidationinhibition) as also the antimicrobial activity (tested towards Gram positive and Gram negative bacteria and eight microfungi) and antiproliferative properties (tested in different human tumor cell lines). Mannitol and trehalose were the main free sugars found in this species (2.01 and 24.71 g/100 g dw, respectively). The fatty acid quantified in higher amounts was linoleic acid (68.00% of total fatty acids). a-Tocopherol was the only isoform of vitam in E detected (55.86 j.lg/1 00 g dw). The organic acids found in this mushroom were oxalic (0.33 g/1 00 g dw), citric (7.97 g/1 00 g dw) and fumaric (0 .1 3 g/100 g dw) acids. p-Hydroxybenzoic acid was the only phenolic acid quantified in C. militaris (0.02 mg/1 00 g dw), but cinnamic acid was also found (0. 11 mg/1 00 g dw). The lowest EC50 values for the antioxidant potential were presented in lipid peroxidation inhibition assays, namely in 13-carotene/l inoleate and TBARS assays (1.05 mg/ml and 0. 77 mg/ml, respectively). The methanolic extract also revealed strong antibacterial and antifungal activities, and it was able to inhibit the proliferation of MCF-7 (breast}, NCI-H460 (non-smalllung),HCT-15 (colon) and Hela (cervical) human carcinoma cell lines. ; FCT (Portugal) and COMPETE/QREN/EU: projects PTOC/AGR-ALI/110062/2009 and PEstOE/AGR/UI0690/2011, and to Serbian Ministry of Education and Science for financial support (grant number 173032).

Abstract Background Mushrooms are not only regarded as gourmet cuisine but also as therapeutic agent to promote cognition health. However, little toxicological information is available regarding their safety. Therefore, the aim of this study was to screen selected ethno-pharmacologically important mushrooms for stimulatory effects on neurite outgrowth and to test for any cytotoxicity. Methods The stimulatory effect of mushrooms on neurite outgrowth was assessed in differentiating mouse neuroblastoma (N2a) cells. Neurite length was measured using Image-Pro Insight processor system. Neuritogenesis activity was further validated by fluorescence immunocytochemical staining of neurofilaments. In vitro cytotoxicity was investigated by using mouse embryonic fibroblast (BALB/3T3) and N2a cells for any embryo- and neuro-toxic effects; respectively. Results Aqueous extracts of Ganoderma lucidum , Lignosus rhinocerotis , Pleurotus giganteus and Grifola frondosa ; as well as an ethanol extract of Cordyceps militaris significantly (p < 0.05) promoted the neurite outgrowth in N2a cells by 38.4 ± 4.2%, 38.1 ± 2.6%, 33.4 ± 4.6%, 33.7 ± 1.5%, and 35.8 ± 3.4%; respectively. The IC 50 values obtained from tetrazolium (MTT), neutral red uptake (NRU) and lactate dehydrogenase (LDH) release assays showed no toxic effects following 24 h exposure of N2a and 3T3 cells to mushroom extracts. Conclusion Our results indicate that G. lucidum , L. rhinocerotis , P. giganteus , G. frondosa and C. militaris may be developed as safe and healthy dietary supplements for brain and cognitive health.

Age-related male sexual dysfunction covers a wide variety of issues, together with spermatogenic and testicular impairment. In the present work, the effects of cordycepin (COR), an active constituent of a nutrient powerhouse Cordyceps militaris Linn, on senile testicular dysfunction in rats was investigated. The sperm kinematics, antioxidant enzymes, spermatogenic factors, sex hormone receptors, histone deacetylating sirtuin 1 (SIRT1), and autophagy-related mammalian target of rapamycin complex 1 (mTORC1) expression in aged rat testes were evaluated. Sprague Dawley rats were divided into young control (2-month-old ; YC), aged control (12-month-old ; AC), and aged plus COR-treated groups (5 (COR-5), 10 (COR-10), and 20 (COR-20) mg/kg). The AC group showed reduced sperm kinematics and altered testicular histomorphology compared with the YC group (p < ; 0.05). However, compared with the AC group, the COR-treated group exhibited improved sperm motility, progressiveness, and average path/straight line velocity (p < ; 0.05&ndash ; 0.01). Alterations in spermatogenesis-related protein and mRNA expression were significantly ameliorated (p < ; 0.05) in the COR-20 group compared with the AC group. The altered histone deacetylating SIRT1 and autophagy-related mTORC1 molecular expression in aged rats were restored in the COR-20 group (p < ; 0.05). In conclusion, the results suggest that COR holds immense nutritional potential and therapeutic value in ameliorating age-related male sexual dysfunctions.

Cordyceps militaris is an entomopathogenic ascomycete with similar pharmacological importance to that of the wild caterpillar fungus Ophiocordyceps sinensis. C. militaris has attracted significant research and commercial interest due to its content in bioactive compounds beneficial to human health and the relative ease of cultivation under laboratory conditions. However, room for improvement exists in the commercial-scale cultivation of C. militaris and concerns issues principally related to appropriate strain selection, genetic degeneration of cultures, and substrate optimization. In particular, culture degeneration—usually expressed by abnormal fruit body formation and reduced sporulation—results in important economic losses and is holding back investors and potential growers (mainly in Western countries) from further developing this highly promising sector. In the present review, the main factors that influence the generation of biomass and metabolites (with emphasis on cordycepin biosynthesis) by C. militaris are presented and evaluated in conjunction with the use of a wide range of supplements or additives towards the enhancement of fungal productivity in large-scale cultivation processes. Moreover, physiological and genetic factors that increase or reduce the manifestation of strain degeneration in C. militaris are outlined. Finally, methodologies for developing protocols to be used in C. militaris functional biology studies are discussed.

As a continuation of our search for biologically active mushroom species the present study investigates in vitro antidiabetic properties of six edible and medicinal mushroom species: Agaricus blazei Murrill, Coprinus comatus (O.F.Müll.) Pers., Cordyceps militaris (L.) Fr., Inonotus obliquus (Ach. ex Pers.) Pilát, Morchella conica Pers. and Phellinus linteus Berk. & M.A. Curtis. In vitro assays on α-amylase and α-glucosidase enzyme inhibition were performed with methanolic extracts of the selected mushrooms. Furthermore, we calculated the necessary daily intake of mushroom extracts and dry mushroom powders based on the equivalent doses of therapeutic drug acarbose given to diabetic patients per day. Our comparative study on enzyme inhibition showed that the most promising potential is ascribed to I. obliquus extract, while no inhibition of α-amylase was recorded with M. conica and C. militaris methanolic extract at the tested concentration. This comparative study is the first highlighting in vitro antidiabetic potential by inhibition of α-amylase and α-glucosidase with methanolic extracts; which makes the investigated speciesmore promising for the diabetes type-2 treatment by an additional and different mechanism of action. ; The authors thank Serbian Ministry of Education, Science and Technological Development for the financial support (grant number 173032). ; info:eu-repo/semantics/publishedVersion

Dietary interventions for people suffering from major depressive disorder (MDD) are an ongoing field of research. In this article, we present a comprehensive background for understanding the possibility of using edible medicinal mushrooms as an adjunctive treatment for MDD. We start with a brief history of MDD, its diagnosis, epidemiology and treatment, and the effects of diet on depression symptoms, followed by a review of neurobiological, behavioral, and clinical studies of medicinal mushrooms. We specifically highlight the results of preclinical and clinical studies on dietary supplementation with three selected mushroom species: Lion's mane (Hericium erinaceus), Caterpillar mushroom (Cordyceps militaris), and Lingzhi/Reishi (Ganoderma lucidum). Preliminary small-sample clinical studies suggest that Lion's mane can influence well-being of humans. In the case of Reishi, the results of clinical studies are equivocal, while in the case of Caterpillar Mushroom, such studies are underway. Edible mushrooms contain 5-hydroxy-L-tryptophan (5-HTP), which is a direct precursor of serotonin—a neurotransmitter targeted in pharmacotherapy of MDD. Therefore, in light of the well-recognized role of stress as a pathogenic factor of MDD, we also describe the neurobiological mechanisms of the interaction between stress and serotonergic neurotransmission; and summarize the current state of knowledge on dietary supplementation with 5-HTP in MDD.

Cordyceps militaris (L.) Link is an important medicinal mushroom used in traditional medicine for treatment of many diseases and improving human health. C. militaris are now widely used in many modern pharmaceutical dosages. It contains many bioactive compounds such as cordycepin, adenosine, sterols, polysaccharides, etc. These compounds provided many beneficial biological activities such as anticancer, antiproliferative, pro-sexual, antioxidant, antibacterial, antifungal, antiviral, immunomodulatory, antiinflammatory and others for C. militaris. The present review highlights the chemical bioactive compounds and pharmacological activities of this medicinal mushroom C. militaris.

BACKGROUND: Due to increasing pressure on natural resources, subsistence agriculture communities in Uganda and Sub-Saharan Africa are experiencing increasingly restricted access to diminishing natural resources that are a critical requirement of their livelihoods. Previously, common-pool resources like forests and grasslands have been either gazetted for conservation or leased for agriculture, the latter in particular for large-scale sugarcane production. Satisfying the increasing consumer demand for grassland or forestry products like wild mushrooms as food or medicine, requires innovative ethno-biological and industry development strategies to improve production capacity, while easing the pressure on diminishing natural resources and averting ecosystems degradation. METHODS: This case study addresses traditional knowledge systems for artisanal mycoculture to identify cultivation practices that enhance sustainable utilization of natural resources. Multi-scalar stakeholder engagement across government and community sectors identified artisanal mushroom producers across five districts in Uganda. Focus groups and semi-structured interviews characterized artisanal production methods and identified locally used substrates for cultivation of different mushroom species. RESULTS: Artisanal practices were characterized for the cultivation of six wild saprophytic mushroom species including Volvariella speciosa (akasukusuku), two Termitomyces sp. (obunegyere and another locally unnamed species), Agaricus sp. (ensyabire) and Agrocybe sp. (emponzira), and one exotic Pleurotus sp. (oyster) that are used as food or medicine. The substrates used for each species differed according to the mushroom's mode of decomposition, those being the following: tertiary decomposers such as those growing under rotting tree stumps or logs from forestry activity like the Agrocybe sp. known as emponzira which grows in forests, thickets, or near homesteads where big logs of hardwood have been left to rot. Also pieces of firewood are chipped off whenever need arises thus providing fuel; secondary decomposers growing on naturally composted grass associated with termites like the Termitomyces sp. known as obunegyere growing in protected sites in gardens, composted cattle manure for Agaricus sp. known as ensyabire in the kraal area where cattle manure is plenty, composted maize cobs for a locally unnamed Agaricus sp. on heaped cobs placed near homesteads; and primary decomposers growing on waste sorghum from brewing the traditional alcoholic drink, muramba for Pleurotus sp. (oyster), and banana and spear grass residue from banana juice processing like the Volvariella speciosa known as akasukusuku because it is associated with the banana plantation locally known in the Luganda language as olusuku and is usually heaped under ficus trees. Management practices also varied based on mode of decomposition and other ecological requirements such as the following: zero tillage and minimal disturbance in areas where obunegyere grow, heaping banana and spear grass residues under the cool ficus trees which also keep them away from banana stump that may cause infestation with nematodes and insects. Even within the generic practices accessibility by the users is critical for example placing logs near homes where children can use them to play, they can be used as fire wood and to even get off-season mushroom as household waste water can make the mushrooms grow. CONCLUSIONS: Our description of artisanal mycoculture methods that respond to conservation and utilization pressures, demonstrates the value of addressing traditional knowledge to improve ethno-biology and mycoculture industry practice. Traditional communities engage in multiple technological and organizational innovations and practices for sustainability and in the case of mushroom production to conserve the environment and culture, ensure variety, food and nutrition security, and income. The results of this study present opportunities to preserve ecosystem quality while developing an artisanal mycoculture system. They have also identified aspects of artisanal mycoculture that most urgently require further ethno-biological study and industry development. Future research and industry development can utilize the result of this study to boost artisanal production of wild saprophytic mushrooms in Sub-Saharan countries, for food or medicinal consumption, and environment conservation. Further development of production efficiencies in context with sustainable natural resource management is recommended.

Mushrooms have bioactive compounds that have antimicrobial, anti-cancer and antioxidant activities among other medicinal benefits. In the present study, we examined the anti-cell proliferation activities of mushrooms from eight genera obtained from the wild in Nigeria. Saccharomyces cerevisiae was used as a model organism to screen mushroom extracts for anti-cell proliferation activity. Polyphenols, high molecular weight polysaccharides and low molecular weight compounds from aqueous extracts were obtained from the test mushrooms using methanol and water respectively. The extracts were screened in vitro at different concentrations of extracts with the CyQuant cell proliferation assay. The high molecular weight polysaccharides from tested mushrooms reduced cell proliferation (96.79% inhibition in Ganoderma multipileum Ding Hou to 66.71% inhibition in Coltricia perennis (L.) Murrill at 10.00mg/ml). Percentage inhibition caused by low molecular weight compounds varied from 94.22% (Ganoderma multipileum) to 76.19% (Coltricia perennis) at 10mg/ml. Percentage of inhibition with the polyphenols varied from 94.12% (Microporus xanthopus Fr) Kuntze) to 79.82% (Coltricia perennis) at high doses. High molecular polysaccharides, low molecular weight compounds and polyphenols from mushrooms have anti-cancer properties. The CyQUANT assay proliferation kit was a very efficient tool for screening extracts from wild mushrooms for anti-cell proliferation activities. Medicinal mushrooms in Nigeria show a lot of promise as a reservoir for drug discovery particularly in the area of cancer research.

The present paper is a review based on published papers to highlight about some potential antiviral mushrooms, which are also found in Nepal. Nepalese mushrooms have not been yet analyzed and investigated so far for their antiviral properties except antioxidant and ACE-Inhibition sources on 62 Nepalese mushrooms as evidence-based complementary and alternative medicine. The published paper from abroad includes the species like: Calvatia gigantia, Cordyceps militaris, Cortinarius caperatus, Fomes fomentarius, Ganoderma lucidum, Grifola frondosa, Hericium erinaceus, Inonotus hispidus, Inonotus obliquus, Lentinula edodes, Omphalotus illudens, Ophiocordyceps sinensis, Phellinus ignarius, Pleurotus citrinopileatus, Pleurotus eryngii. Pleurotus ostreatus, Polyporus umbellatus, Russula delica. Schizophyllum commune, Scleroderma citrinum, Tremella sp. and Trametes versicolor, which are also found in the natural temperate (Schima – Castanopsis - Rhododrendron – Quercus) forests of Kathmandu valley. This paper may provide the researchers a glimpse to the aspects of experiment and lead to investigate on medicinal, antiviral and antibacterial properties of mushrooms found in Nepal. Gautam & Adhikari (2007) have screened Ophicordyceps sinensis for its chemical compound.

Phytohormones cytokinins are known to promote cell division in plants. Contrary, in animal's and human's tissues they induce apoptosis and block the cell cycle of a wide spectrum of tumour cells. Therapeutic effects of cytokinins, specifically their anticancer and immunomodulatory actions are similar to those of medicinal mushrooms. We detected cytokinins in mycelial biomass of two species of medicinal mushrooms growing in vitro (Fomitopsis officinalis strain 5004 and Hericium coralloides strain 2332) using HPLC-MS. Trans-zeatin, zeatin riboside, zeatin-O-glucoside and isopentenyladenine were found. Crude extracts and purified cytokinin fractions from mycelial biomass were tested on the growth and development of cultures of tumor cells lines: Hela (MTT-assay), T24/83 (viability and level apoptotic cells) and HepG2 (consumption of glucose). The effect of cytokinin fraction from mycelial biomass of Fomitopsis officinalis on pathogenic cells was higher compared to Hericium coralloides one. The data obtained revealed a higher cytotoxic/cytostatic effect of the purified cytokinin fractions in comparison with crude methanolic extracts; also higher apoptotic index was found. Under the influence of the test agents the intensification of glucose uptake into cells was observed. This indicator was higher for crude mushroom mycelium extracts, whereas under the action of purified fractions the glucose uptake rate was lower, thus decreased glycolysis level was recorded. Also, the effect of both crude extract and purified fraction from H. coralloides mycelial biomass on glucose uptake in the conditioned medium was lower against F. officinalis.These results confirm the assumption that biologically active substances of medicinal mushrooms with high pharmacological potential include cytokinins.

Diabetes mellitus is a chronic disorder of metabolism followed by abnormal rise in plasma glucose levels, as a consequence of unequilibrated insulin production and/or insensitivity to the effect of this hormone in signal transduction of cellular receptors. One of the effective strategies for diabetes type-2 management is the inhibition of complex polysaccharide hydrolysis by pancreatic α-amylase and absorption limitation of glucose by inhibiting intestinal α-glucosidase enzyme. Agaricus blazei Murrill, Coprinus comatus (O.F.Müll.) Pers., Cordyceps militaris (L.) Fr., Inonotus obliquus (Ach. ex Pers.) Pilát, Morchella conica Pers. and Phellinus linteus Berk. & M.A. Curtis were investigated for their antidiabetic properties. In vitro assays on α-amylase and α-glucosidase enzyme inhibition were performed with methanolic extracts of the selected mushrooms. Furthermore, we calculated the necessary daily intake of mushroom extracts and dry mushroom powders based on the equivalent doses of therapeutic drug acarbose given to diabetic patients per day. Our comparative study on enzyme inhibition showed that the most promising potential is ascribed to I. obliquus extract, while no inhibition of α-amylase was recorded with M. conica and C. militaris methanolic extract at the tested concentration. The lowest daily intake of mushroom powder was suggested for I. obliquus with the dose of 3 × 1.148 g/day, while the highest was predicted for P. linteus 3 × 2.215 g/day. Although majority of previous studies showed in vivo antidiabetic potential of water and polysaccharidic mushroom extracts by different experimental approaches, our study is the first highlighting in vitro antidiabetic potential by inhibition of α-amylase and α-glucosidase with methanolic extracts; which makes the investigated species more promising for the diabetes type-2 treatment by another additional and different mechanism of action. ; Soković M, editor. Book of abstracts: The 2nd Balkans-China Mini-symposium on Natural Products and Drug Discovery; 2019 Apr 11-13; Belgrade, Serbia. Belgrade: Institute for Biological Research "Siniša Stanković", University of Belgrade; 2019. p 37.