Spent substrate from mushroom cultivation: exploitation potential toward various applications and value-added products.

Spent mushroom substrate (SMS) is the residual biomass generated after harvesting the fruitbodies of edible/medicinal fungi. Disposal of SMS, the main by-product of the mushroom cultivation process, often leads to serious environmental problems and is financially demanding. Efficient recycling and valorization of SMS are crucial for the sustainable development of the mushroom industry in the frame of the circular economy principles. The physical properties and chemical composition of SMS are a solid fundament for developing several applications, and recent literature shows an increasing research interest in exploiting that inherent potential. This review provides a thorough outlook on SMS exploitation possibilities and discusses critically recent findings related to specific applications in plant and mushroom cultivation, animal husbandry, and recovery of enzymes and bioactive compounds.

Valorization of spent substrate is crucial for a sustainable mushroom industry.The review covers spent mushroom substrate (SMS) valorization for multiple uses.SMS composition and mushroom species are essential factors for its exploitation.SMS valorization forms an integral part of cascade use of plant biomass.

Correction: Boulaka et al. Genoprotective Properties and Metabolites of ß-Glucan-Rich Edible Mushrooms Following Their In Vitro Fermentation by Human Faecal Microbiota. Molecules 2020, 25, 3554.

The authors wish to add the following information to the Authors and Affiliation section of our paper published in Molecules [...].

Evaluation of the lignocellulose degradation potential of Mediterranean forests soil microbial communities through diversity and targeted functional metagenomics.

The enzymatic arsenal of several soil microorganisms renders them particularly suitable for the degradation of lignocellulose, a process of distinct ecological significance with promising biotechnological implications. In this study, we investigated the spatiotemporal diversity and distribution of bacteria and fungi with 16S and Internally Trascribed Spacer (ITS) ribosomal RNA next-generation-sequencing (NGS), focusing on forest mainland Abies cephalonica and insular Quercus ilex habitats of Greece. We analyzed samples during winter and summer periods, from different soil depths, and we applied optimized and combined targeted meta-omics approaches aiming at the peroxidase-catalase family enzymes to gain insights into the lignocellulose degradation process at the soil microbial community level. The microbial communities recorded showed distinct patterns of response to season, soil depth and vegetation type. Overall, in both forests Proteobacteria, Actinobacteria, Acidobacteria were the most abundant bacteria phyla, while the other phyla and the super-kingdom of Archaea were detected in very low numbers. Members of the orders Agaricales, Russulales, Sebacinales, Gomphales, Geastrales, Hysterangiales, Thelephorales, and Trechisporales (Basidiomycota), and Pezizales, Sordariales, Eurotiales, Pleosporales, Helotiales, and Diaporthales (Ascomycota) were the most abundant for Fungi. By using optimized "universal" PCR primers that targeted the peroxidase-catalase enzyme family, we identified several known and novel sequences from various Basidiomycota, even from taxa appearing at low abundance. The majority of the sequences recovered were manganese peroxidases from several genera of Agaricales, Hysterangiales, Gomphales, Geastrales, Russulales, Hymenochaetales, and Trechisporales, while lignin -and versatile-peroxidases were limited to two to eight species, respectively. Comparisons of the obtained sequences with publicly available data allowed a detailed structural analysis of polymorphisms and functionally relevant amino-acid residues at phylogenetic level. The targeted metagenomics applied here revealed an important role in lignocellulose degradation of hitherto understudied orders of Basidiomycota, such as the Hysterangiales and Gomphales, while it also suggested the auxiliary activity of particular members of Proteobacteria, Actinobacteria, Acidobacteria, Verrucomicrobia, and Gemmatimonadetes. The application of NGS-based metagenomics approaches allows a better understanding of the complex process of lignocellulolysis at the microbial community level as well as the identification of candidate taxa and genes for targeted functional investigations and genetic modifications.

In Vitro Fermentation of Edible Mushrooms: Effects on Faecal Microbiota Characteristics of Autistic and Neurotypical Children.

Children with autism spectrum disorder (ASD) often suffer gastrointestinal disturbances consistent with gut microbiota (GM) alterations. Treatment with pro/prebiotics may potentially alleviate gut symptoms, but the evidence for prebiotics is scarce. This study aims to evaluate the effects of edible mushrooms (Pleurotus, Basidiomycota) and prebiotic compounds on GM composition and metabolite production in vitro, using faecal samples from autistic and non-autistic children. Specific microbial populations were enumerated after 24 h of fermentation by quantitative PCR, and the metabolic production was determined by gas chromatography. Higher levels of Prevotella spp. and Bifidobacterium spp. were measured in neurotypical children compared to ASD children. A total of 24 h fermentation of Pleurotus eryngii and P. ostreatus mushroom powder increased the levels of Bifidobacterium, while known prebiotics increased the levels of total bacteria and Bacteroides in both groups. Only P. eryngii mushrooms resulted in significantly elevated levels of total bacteria Bacteroides and Feacalibacterium prausnitzii compared to the negative control (NC) in the ASD group. Both mushrooms induced elevated levels of butyrate after 24 h of fermentation, while short-chain fructooligosaccharides induced increased levels of acetate in the ASD group, compared to NC. Overall, this study highlights the positive effect of edible mushrooms on the GM and metabolic activity of children with ASD.

Pleurotus eryngii Chips-Chemical Characterization and Nutritional Value of an Innovative Healthy Snack.

Nowadays, as the pandemic has reshaped snacking behaviors, and consumers have become more health-conscious, the need for the incorporation of "healthy snacking" in our diets has emerged. Although there is no agreed-upon definition of "healthy snacking", dietary guidelines refer to snack foods with high nutritional and biological value. The aim of this study was to chemically characterize and determine the nutritional value of an innovative UVB-irradiated and baked snack from Pleurotus eryngii mushrooms. P. eryngii is an edible mushroom native to the Mediterranean basin. We applied proximate composition, amino acids, fatty acids, vitamins, and macro and trace elements analyses. Also, we computed indices to assess the nutritional quality of food, and we evaluated the sensory characteristics of the mushroom snack. We found high nutritional, consumer, and biological values for the snack. More specifically it was low in calories, high in fibre and protein, low in lipids, without added sugars, and high in ergosterol and beta-glucans. Additionally, it had some vitamins and trace elements in significant quantities. Its NRF9.3 score was considerably high compared to most popular snacks, and the snack exhibited high hypocholesterolemic and low atherogenic and thrombogenic potentials. In conclusion, as a result of UVB-irradiation and baking of P. eryngii mushrooms, the snack's nutritional and biological value were not affected; instead, it provided a "healthy snacking" option.

In Vitro Fermentation of Pleurotus eryngii Mushrooms by Human Fecal Microbiota: Metataxonomic Analysis and Metabolomic Profiling of Fermentation Products.

Edible mushrooms contain biologically active compounds with antioxidant, antimicrobial, immunomodulatory and anticancer properties. The link between their anticancer and immunomodulatory properties with their possible prebiotic activity on gut micro-organisms has been the subject of intense research over the last decade. Lyophilized Pleurotus eryngii (PE) mushrooms, selected due to their strong lactogenic effect and anti-genotoxic, immunomodulatory properties, underwent in vitro static batch fermentation for 24 h by fecal microbiota from eight elderly apparently healthy volunteers (>65 years old). The fermentation-induced changes in fecal microbiota communities were examined using Next Generation Sequencing of the hypervariable regions of the 16S rRNA gene. Primary processing and analysis were conducted using the Ion Reporter Suite. Changes in the global metabolic profile were assessed by 1H NMR spectroscopy, and metabolites were assigned by 2D NMR spectroscopy and the MetaboMiner platform. PLS-DA analysis of both metataxonomic and metabolomic data showed a significant cluster separation of PE fermented samples relative to controls. DEseq2 analysis showed that the abundance of families such as Lactobacillaceae and Bifidobacteriaceae were increased in PE samples. Accordingly, in metabolomics, more than twenty metabolites including SCFAs, essential amino acids, and neurotransmitters discriminate PE samples from the respective controls, further validating the metataxonomic findings.

Investigation of Abortiporus biennis lignocellulolytic toolbox, and the role of laccases in polystyrene degradation.

White-rot basidiomycetes are the only microorganisms able to produce both hydrolytic (cellulases and hemicellulases) and oxidative (ligninolytic) enzymes for degrading all lignocellulose constituents. Their enzymatic machinery makes them ideal for the discovery of novel enzymes with desirable properties. In the present work, Abortiporus biennis, a white-rot fungus, was studied in regard to its lignocellulolytic potential. Secretomics and biochemical analyses were employed to study the strain's enzymatic arsenal, after growth in corn stover cultures and xylose-based defined media. The results revealed the presence of all the necessary enzymatic activities for complete breakdown of biomass, while the prominent role of oxidative enzymes in the lignocellulolytic strategy of the strain became evident. Two novel laccases, AbiLac1 and AbiLac2, were isolated from the culture supernatant with ion-exchange chromatography. Characterization of purified laccases revealed their ability to oxidize a wide variety of phenolic and non-phenolic substrates. AbiLac1 was found to oxidize polystyrene powder, showing high depolymerization potential, based on radical chain scission mechanism as evidenced by molecular weight decrease. The results of the present study demonstrate the biotechnological potential of the unexplored enzymatic machinery of white-rot basidiomycetes, including the design of improved lignocellulolytic cocktails, as well as the degradation and/or valorization of plastic waste materials.

A Randomized Controlled Trial on Pleurotus eryngii Mushrooms with Antioxidant Compounds and Vitamin D2 in Managing Metabolic Disorders.

This study examined the effects of a Pleurotus eryngii mushroom snack on metabolically unhealthy patients. After harvest, mushrooms were baked and subjected to UV-B irradiation to enhance vitamin D2 content. A randomized controlled trial was conducted for three months with two arms. Both groups received conventional nutritional counseling for metabolic disorders, while the intervention group had to consume the snack daily as well. We collected blood samples at the beginning and the end of the study to determine biochemical measurements and serum 25(OH)D2 and to evaluate inflammation and oxidative stress. One hundred patients consented and were randomized. Comparatively to the control group, snack consumption regulated glucose levels and reduced body weight, fat, waist and hip circumferences. In addition, 25(OH)D2 increased significantly in the intervention group. The levels of LDL and SGOT were lower only in the intervention group. Levels of IL-6 and ox-LDL decreased in the mushroom group, while the overall physical health increased. These findings suggest potential antidiabetic, antiobesity, anti-inflammatory and antioxidant health benefits of the snack to metabolically unhealthy individuals.

Genoprotective activity of the Pleurotus eryngii mushrooms following their in vitro and in vivo fermentation by fecal microbiota.

Pleurotus eryngii mushrooms are commercially cultivated and widely consumed due to their organoleptic properties, and the low caloric and high nutritional value. In addition, they contain various biologically active and health-promoting compounds; very recently, their genoprotective effect in Caco-2 cells after their fermentation by the human fecal microbiota was also documented. In the current study, the effect of P. eryngii pre- and post-fermentation supernatants in micronuclei formation was evaluated in human lymphocytes. In addition, the genoprotective properties of increasing concentrations of aqueous extracts from P. eryngii mushrooms (150, 300, 600 mg/kg) against the cyclophosphamide-induced DNA damage were studied in young and elderly female and male mice in bone marrow and whole blood cells. The ability of the highest dose (600 mg/kg) to regulate the main cellular signaling pathways was also evaluated in gut and liver tissues of female animals by quantifying the mRNA expression of NrF2, Nfkß, DNMT1, and IL-22 genes. P. eryngii post-fermentation, but not pre-fermentation, supernatants were able to protect human lymphocytes from the mitomycin C-induced DNA damage in a dose-dependent manner. Similarly, genoprotection was also observed in bone marrow cells of mice treated by gavage with P. eryngii extract. The effect was observed in all the experimental groups of mice (young and elderly, male and female) and was more potent in young female mice. Overexpression of all genes examined was observed in both tissues, mainly among the elderly animals. In conclusion, P. eryngii mushrooms were shown to maintain genome integrity through protecting cells from genotoxic insults. These beneficial effects can be attributed to their antioxidant and immunomodulatory properties, as well as their ability to regulate the cell's epigenetic mechanisms and maintain cell homeostasis.

Immunomodulating Activity of Pleurotus eryngii Mushrooms Following Their In Vitro Fermentation by Human Fecal Microbiota.

Recent studies have revealed the crucial role of several edible mushrooms and fungal compounds, mainly polysaccharides, in human health and disease. The investigation of the immunomodulating effects of mushroom polysaccharides, especially ß-glucans, and the link between their anticancer and immunomodulatory properties with their possible prebiotic activity on gut micro-organisms has been the subject of intense research over the last decade. We investigated the immunomodulating effects of Pleurotus eryngii mushrooms, selected due to their high ß-glucan content, strong lactogenic effect, and potent geno-protective properties, following in vitro fermentation by fecal inocula from healthy elderly volunteers (>60 years old). The immunomodulating properties of the fermentation supernatants (FSs) were initially investigated in U937-derived human macrophages. Gene expression as well as pro- (TNF-α, IL-1ß) and anti-inflammatory cytokines (IL-10, IL-1Rα) were assessed and correlated with the fermentation process. The presence of P. eryngii in the fermentation process led to modifications in immune response, as indicated by the altered gene expression and levels of the cytokines examined, a finding consistent for all volunteers. The FSs immunomodulating effect on the volunteers' peripheral blood mononuclear cells (PBMCs) was verified through the use of cytometry by time of flight (CyTOF) analysis.

Enhancing the nutritional and functional properties of Pleurotus citrinopileatus mushrooms through the exploitation of winery and olive mill wastes.

Treatment and disposal of wineries and olive-oil mills waste is usually associated with complex processes, which are often of limited wide-scale applicability. Olive-leaves plus two-phase olive mill waste (OLW) or grape marc plus wheat straw (GMW) were assessed as substrates for the cultivation of the choice edible mushroom Pleurotus citrinopileatus. GMW led to increased mushroom biological efficiency and shorter production cycles. Antioxidant activities, triterpenic acids, free amino acids, lovastatin and ergosterol were significantly higher in fruitbodies from GMW; the latter compound was positively correlated with squalene concentrations in substrates. Glucans, resveratrol and fatty acids content showed minor differences among mushrooms from the three substrates examined, whereas ergothioneine was significantly higher in fruitbodies grown on OLW. High correlations were noted for oleanolic, ursolic and amino acid content in mushrooms and their respective substrates. Moreover, FTIR spectra revealed variations in fruitbodies content in bioactive compounds which were associated with the substrates used.

Pleurotus eryngii improves postprandial glycaemia, hunger and fullness perception, and enhances ghrelin suppression in people with metabolically unhealthy obesity.

The aim of this study was to examine potential postprandial benefits of Pleurotus eryngii in nineteen volunteers with metabolically unhealthy obesity. An acute, randomized, crossover-designed trial comparing a meal with Pleurotus eryngii and a control meal was performed. The two meals matched in macronutrient and caloric content. Participants consumed both meals in random order after an overnight fast. Blood samples were drawn before and 30, 60, 90, 120, 150 and 180 min after meal consumption (in total 266 samples) to determine glucose, insulin, ghrelin, peptide YY, glucagon-like peptide-1 and glicentin. Visual analog scales measuring the subjective perception of hunger and fullness were completed at the same time points. The test meal resulted in lower glucose incremental area under the curve (iAUC). Additionally, the iAUC of the ghrelin response over time was substantially lower after the test meal (p = 0.033). Lower desire to eat and higher fullness was reflected by significantly lower hunger iAUC (p = 0.046) and higher fullness iAUC (p = 0.042) after the test meal. No differences in insulin, PYY, GLP-1 and glicentin were observed. Pleurotus eryngii can ameliorate postprandial glycaemia, appetite and regulate ghrelin levels at the postprandial state. This effect is attributed to the bioactive polysaccharides that inhibit the activity of enzymes catalysing carbohydrate hydrolysis, cause a delayed gastric emptying and glucose absorption.

Biomass and Cordycepin Production by the Medicinal Mushroom Cordyceps militaris-A Review of Various Aspects and Recent Trends towards the Exploitation of a Valuable Fungus.

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.

Fermentation Supernatants of Pleurotus eryngii Mushroom Ameliorate Intestinal Epithelial Barrier Dysfunction in Lipopolysaccharide-Induced Caco-2 Cells via Upregulation of Tight Junctions.

In recent years, modulation of gut microbiota through prebiotics has garnered interest as a potential to ameliorate intestinal barrier dysfunction. The aim of the study was to examine the in vitro effect of fermentation supernatants (FSs) from rich in ß-glucan Pleurotus eryngii mushrooms on the expression levels of tight junctions (TJs) genes in Caco-2 cells stimulated by bacterial lipopolysaccharides (LPS). Mushrooms were fermented using fecal inocula in an in vitro batch culture model. Caco-2 cells were subjected to LPS and FS treatment under three different conditions: pre-incubation with FS, co- and post-incubation. Reverse transcription PCR was applied to measure the expression levels of zonulin-1, occludin and claudin-1 genes. FSs from P. eryngii mushrooms led to a significant upregulation of the TJs gene expression in pre-incubation state, indicating potential preventive action. Down-regulation of all TJs gene expression levels was observed when the cells were challenged with LPS. The FS negative control (gut microbiota of each donor with no carbohydrate source) exhibited a significant upregulation of TJs expression levels compared to the cells that were challenged with LPS, for all three conditions. Overall, our data highlighted the positive and potential protective effects of P. eryngii mushrooms in upregulation of TJs' genes.

Volatile Profiling of Pleurotus eryngii and Pleurotus ostreatus Mushrooms Cultivated on Agricultural and Agro-Industrial By-Products.

The influence of genetic (species, strain) and environmental (substrate) factors on the volatile profiles of eight strains of Pleurotus eryngii and P. ostreatus mushrooms cultivated on wheat straw or substrates enriched with winery or olive oil by products was investigated by headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS). Selected samples were additionally roasted. More than 50 compounds were determined in fresh mushroom samples, with P. ostreatus presenting higher concentrations but a lower number of volatile compounds compared to P. eryngii. Roasting resulted in partial elimination of volatiles and the formation of pyrazines, Strecker aldehydes and sulfur compounds. Principal component analysis on the data obtained succeeded to discriminate among raw and cooked mushrooms as well as among Pleurotus species and strains, but not among different cultivation substrates. Ketones, alcohols and toluene were mainly responsible for discriminating among P. ostreatus strains while aldehydes and fatty acid methyl esters contributed more at separating P. eryngii strains.

Mallocybe heimii ectomycorrhizae with Cistus creticus and Pinus halepensis in Mediterranean littoral sand dunes - assessment of phylogenetic relationships to M. arenaria and M. agardhii.

Ectomycorrhizal symbiosis appears extensively in the Northern Hemisphere, where Mediterranean ecosystems constitute an important ecological area of considerable biodiversity value. Littoral sand dunes are among high-risk habitats, and ectomycorrhizal lifestyle contributes significantly to supporting life in such regions. Mallocybe heimii (Bon) Matheny & Esteve-Rav. (Inocybaceae, Basidiomycota) and the very similar M. arenaria (Bon) Matheny & Esteve-Rav. grow in poor, usually sandy soils, in association with angiosperms or gymnosperms. Basidiomata originally identified under these names were collected from littoral sand dunes of Greece, and their morpho-anatomical characteristics were examined in conjunction with material derived from other European regions. Sequences from basidiomata and root tips corresponding to the nuclear rDNA internal transcribed spacer region (ITS) and large subunit (LSU) were obtained and analyzed. Phylogenetic results demonstrated that material identified as M. heimii or M. arenaria form a single well-supported group, while M. agardhii (N. Lund) Matheny & Esteve-Rav. is confirmed to be distinct from M. arenaria (the latter was initially described as a variety of the former, i.e., I. agardhii var. arenaria Bon). A detailed tree of the genus Mallocybe was generated on the basis of concatenated ITS and LSU sequences, and relationships of selected taxa are discussed in the light of morphological and sequence data. In addition, the first morphotype descriptions of M. heimii ectomycorrhizae with Cistus creticus L. and Pinus halepensis Miller are hereby provided. Both morphotypes exhibited the typical characteristics of Inocybe/Mallocybe ectomycorrhizae; however, differences were noted, the most significant being the presence of clamps on mantle hyphae and the type of anastomoses.

Effects of fungal beta-glucans on health - a systematic review of randomized controlled trials.

INTRODUCTION: Beta-glucans are polysaccharides that exhibit a wide range of biological properties as a result of their varying chemical composition. Like all dietary fibers, they avoid catabolism in the upper gastrointestinal tract, and they reach the large intestine undigested. There, they undergo fermentation by the gut microbiota, a process that has potential beneficial effects for the host. The aim of this systematic review is to assess the effects of consumption of beta-(1 → 3,1 → 6)-d-glucans, naturally found in the cell walls of fungi, on health outcomes. METHODS: A comprehensive literature search was performed on PubMed, Cochrane Library and Web of Science to retrieve studies that applied randomized controlled trials (RCTs) to investigate the impact of exclusive oral administration of fungal beta-glucans in any form and at any dosage to healthy subjects or patients. RESULTS: Thirty-four RCTs, of the 917 records retrieved in total, met the eligibility criteria and are included in the present review. The sources of fungal beta-glucans were Saccharomyces cerevisiae, Aureobasidium pullulans, Pleurotus ostreatus, Lentinula edodes and Ganoderma lucidum, and the dosage of supplementation ranged from 2.5 to 1000 mg daily for up to 6.5 months. The primary physiological outcome of the majority of the interventions was immunomodulation, which resulted in (a) strengthened immune defense that reduces the incidence and symptoms of cold, flu and other respiratory infections and (b) improvement of allergic symptoms. However, the findings on the induction of immune response alterations were inconsistent at the cellular and molecular levels. Another aspect is psychological wellbeing, as the cohorts that received the polysaccharides of interest reported improvement in their mood states as well as amelioration of overall wellbeing. At the same time, it might also be useful as a complementary agent to patients undergoing cancer therapies. Furthermore, supplements containing beta-(1 → 3,1 → 6)-d-glucan administered to overweight/obese adults might have the potential to decrease comorbid conditions associated with obesity. Notably, no adverse event causally related to glucans was recorded. CONCLUSIONS: Supplementation with beta-(1 → 3,1 → 6)-d-glucans is well-tolerated, and health-promoting properties are manifested primarily through the potentiation of the immune system. More studies are required to confirm their additional beneficial effects, to establish the optimal dose, and to reveal the underlying molecular mechanisms.

On the Identification and Quantification of Ergothioneine and Lovastatin in Various Mushroom Species: Assets and Challenges of Different Analytical Approaches.

In recent years, mushrooms have drawn the attention of agro-industries and food-industries as they were considered to be valuable natural sources of health promoting compounds such as ß-glucans, ergothioneine, and lovastatin. The detection and quantification of such compounds by implementing reliable analytical approaches is of the utmost importance in order to adjust mushrooms' cultivation conditions and maximize the production in different species. Toward this direction, the current study focuses on the comparison of ultraviolet-visible (UV-Vis) spectrometry and liquid chromatography-mass spectrometry (LC-MS) methods (a) by evaluating the content of ergothioneine and lovastatin in mushrooms and (b) by highlighting any possible substrate-based interferences that hinder the accurate determination of these two compounds in order to propose the technique-of-choice for a standardized bioactive compounds monitoring. For this purpose, mushrooms produced by three species (i.e., Agaricus bisporus, Pleurotus ostreatus, and P. citrinopileatus) on various cultivation substrates, namely wheat straw (WS), winery (grape marc (GM)), and olive oil (OL) by-products, were examined. Among the two applied techniques, the developed and validated LC-MS methods, exhibiting relatively short analysis time and higher resolution, emerge as the methods-of-choice for detecting ergothioneine and lovastatin in mushrooms. On the contrary, UV-Vis methods were hindered due to co-absorbance of different constituents, resulting in invalid results. Among the studied mushrooms, P. citrinopileatus contained the highest amount of ergothioneine (822.1 ± 20.6 mg kg-1 dry sample), whereas A. bisporus contained the highest amounts of lovastatin (1.39 ± 0.014 mg kg-1 dry sample). Regarding the effect of different cultivation substrates, mushrooms produced on OL and WS contained the highest amount of ergothioneine, while mushrooms deriving from GM-based substrates contained the highest amount of lovastatin.

Fermentation of Pleurotus ostreatus and Ganoderma lucidum mushrooms and their extracts by the gut microbiota of healthy and osteopenic women: potential prebiotic effect and impact of mushroom fermentation products on human osteoblasts.

Recent data have highlighted the role of the gut microbiota and its several metabolites in maintaining bone health. Thus, gut microbiota manipulation, e.g., by prebiotics, might offer a plausible target in the fight against bone degenerative diseases. This study aimed (a) to investigate the in vitro prebiotic potential of Ganoderma lucidum and Pleurotus ostreatus mushrooms in healthy and osteopenic women and (b) to explore the impact of mushroom fermentation products on human osteoblasts. G. lucidum LGAM 9720 and P. ostreatus IK 1123 lyophilized mushroom-powders (2% w/v) and their hot-water extracts (1% w/v) were fermented in a 24 h static batch culture model by using faecal inocula from healthy (n = 3) or osteopenic (n = 3) donors. Gut microbiota analysis (qPCR) and measurement of short chain fatty acids (SCFAs) were performed during fermentation, and 24 h-prebiotic indexes were calculated. Evaluation of the effects of fermentation products on bone metabolism parameters (OPG: osteoprotegerin; and RANKL: receptor activator of nuclear factor kappa B ligand) in osteoblast cultures was also performed. Our data suggest that the origin of the gut microbiota inoculum plays a major role in the viability of osteoblasts. The treatments using P. ostreatus mushroom-powder and G. lucidum mushroom-extract had positive effects based on gut microbiota and SCFA analyses. Both mushrooms exhibited lower RANKL levels compared to controls, whereas their extracts tended to enhance the osteoblastic activity. In conclusion, mushrooms that are rich in beta-glucans may exert beneficial in vitro effects on bone physiology by alterations in the gut microbiota and/or SCFA production.

A global meta-analysis of ITS rDNA sequences from material belonging to the genus Ganoderma (Basidiomycota, Polyporales) including new data from selected taxa.

Ganoderma P. Karst. is a cosmopolitan genus of white-rot fungi which comprises species with highly-prized pharmaceutical properties, valuable biotechnological applications and of significant phytopathological interest. However, the status of the taxonomy within the genus is still highly controversial and ambiguous despite the progress made through molecular approaches. A metadata analysis of 3908 nuclear ribosomal internal transcribed spacer (ITS) rDNA sequences obtained from GenBank/ENA/DDBJ and UNITE was performed by targeting sequences annotated as Ganoderma, but also sequences from environmental samples and from material examined for the first time. Ganoderma taxa segregated into five main lineages (Clades A to E). Clade A corresponds to the core of laccate species and includes G. shanxiense and three major well-supported clusters: Cluster A.1 ('G. lucidum sensu lato') consists of taxa from Eurasia and North America, Cluster A.2 of material with worldwide occurrence including G. resinaceum and Cluster A.3 is composed of species originating from all continents except Europe and comprises G. lingzhi. Clade B includes G. applanatum and allied species with a Holarctic distribution. Clade C comprises taxa from Asia and Africa only. Clade D consists of laccate taxa with tropical/subtropical occurrence, while clade E harbours the highest number of non-laccate species with a cosmopolitan distribution. The 92 Ganoderma-associated names, initially used for sequences labelling, correspond to at least 80 taxa. Amongst them, 21 constitute putatively new phylospecies after our application of criteria relevant to the robustness/support of the terminal clades, intra- and interspecific genetic divergence and available biogeographic data. Moreover, several other groups or individual sequences seem to represent distinct taxonomic entities and merit further investigation. A particularly large number of the public sequences was revealed to be insufficiently and/or incorrectly identified, for example, 87% and 78% of entries labelled as G. australe and G. lucidum, respectively. In general, ITS demonstrated high efficacy in resolving relationships amongst most of the Ganoderma taxa; however, it was not equally useful at elucidating species barriers across the entire genus and such cases are outlined. Furthermore, we draw conclusions on biogeography by evaluating species occurrence on a global scale in conjunction with phylogenetic structure/patterns. The sequence variability assessed in ITS spacers could be further exploited for diagnostic purposes.