Macrofungal Extracts as a Source of Bioactive Compounds for Cosmetical Anti-Aging Therapy: A Comprehensive Review.

For centuries, mushrooms have been used as a component of skincare formulations. Environmental stresses and a modern lifestyle expose the skin to accelerated aging. To slow down this process, natural anti-aging skincare ingredients are being sought. In this review, 52 scientific publications about the effects of chemical compounds extracted from the fruiting bodies of macrofungi on skin cells were selected. The effects of extracts from nine species that are tested for anti-aging effects have been described. According to available literature data, macrofungi contain many polysaccharides, phenolic compounds, polysaccharide peptides, free amino acids, sterols, proteins, glycosides, triterpenes, alkaloids, which can have an anti-aging effect on the skin by acting as antioxidants, photoprotective, skin whitening, moisturizing, anti-inflammatory and stabilizing collagen, elastin and hyaluronic acid levels in the skin.

Agaricus Subrufescens ameliorates ovarian dysfunction and regulates altered biochemical parameters in rats with Letrozole induced polycystic ovarian syndrome.

OBJECTIVE: The objective of this study is to investigate the effects of an ethanolic extract derived from Agaricus subrufescens on rat models exhibiting Polycystic Ovarian Syndrome (PCOS) induced by Letrozole. METHODS: A total of thirty female Wistar rats were divided into five groups, each consisting of six rats. The negative control group was administered a volume of 1 mL of a 0.5% solution of carboxy methylcellulose (CMC). Letrozole (1 mg/kg) was administered to additional groups for a duration of 21 days in order to induce polycystic ovary syndrome (PCOS). Animals designated as positive controls were euthanized on the 22nd day. Both the test group and the standard group were subjected to treatment from the 22nd day to the 36th day. The experimental group was administered ethanolic extract of Agaricus subrufescens at doses of 200 mg/kg and 400 mg/kg p.o, while the control group received clomiphene citrate at a dose of 1 mg/kg. The study observed various physiological markers in individuals with polycystic ovarian disease, including estimated blood glucose levels, total cholesterol levels, triglyceride levels, and hormonal fluctuations such as increased testosterone and estrogen levels, as well as decreased progesterone levels. The presence of menstrual irregularities was confirmed through the examination of vaginal smears and histopathological changes in the ovaries. RESULTS: The consumption of Agaricus subrufescens was found to have a significant impact on various physiological parameters, including blood glucose levels, testosterone levels, anovulation, and menstrual irregularity. All therapeutic interventions significantly normalized the levels of serum glutamic-oxaloacetic transaminase (SGOT) and serum glutamic-pyruvic transaminase (SGPT). The rats with polycystic ovary syndrome (PCOS) that were induced by Letrozole exhibited increased levels of urea and creatinine. The findings of this study indicate that the administration of Agaricus subrufescens therapy has a protective effect on renal function, as evidenced by a reduction in serum levels of urea and creatinine. In rats with polycystic ovary syndrome (PCOS) induced by Letrozole, the inhibition of hepatic synthesis, promotion of ovarian follicle immaturity, and elevation of androgen secretions result in an increase in the weight of the liver and ovaries. The weight of endocrine organs exhibited a decrease across all treatment groups. The histopathological examination of PCOS specimens revealed an increased presence of cysts and theca lutein cells. The group of rats with polycystic ovary syndrome (PCOS) that did not receive treatment exhibited a higher number of cysts compared to the groups that received treatment. CONCLUSION: This study demonstrated that the administration of Letrozole orally resulted in the development of polycystic ovarian disease. The results indicated heightened levels of blood glucose, total cholesterol, and triglycerides, as well as alterations in hormone levels such as increased testosterone and estrogen, and decreased progesterone. These hormonal changes were accompanied by menstrual irregularities, which were confirmed through the examination of vaginal smears and histopathological analysis of the ovaries in the control group with polycystic ovarian disease. The treatment groups that received Agaricus subrufescens exhibited a decrease in blood glucose, total cholesterol, and testosterone levels.

Decoding the chromosome-scale genome of the nutrient-rich Agaricus subrufescens: a resource for fungal biology and biotechnology.

Agaricus subrufescens, also known as the "sun mushroom," has significant nutritional and medicinal value. However, its short shelf life due to the browning process results in post-harvest losses unless it's quickly dehydrated. This restricts its availability to consumers in the form of capsules. A genome sequence of A. subrufescens may lead to new cultivation alternatives or the application of gene editing strategies to delay the browning process. We assembled a chromosome-scale genome using a hybrid approach combining Illumina and Nanopore sequencing. The genome was assembled into 13 chromosomes and 31 unplaced scaffolds, totaling 44.5 Mb with 96.5% completeness and 47.24% GC content. 14,332 protein-coding genes were identified, with 64.6% of the genome covered by genes and 23.41% transposable elements. The mitogenome was circularized and encoded fourteen typical mitochondrial genes. Four polyphenol oxidase (PPO) genes and the Mating-type locus were identified. Phylogenomic analysis supports the placement of A. subrufescens in the Agaricomycetes clade. This is the first available genome sequence of a strain of the "sun mushroom." Results are available through a Genome Browser (https://plantgenomics.ncc.unesp.br/gen.php?id=Asub) and can support further fungal biological and genomic studies.

Agaricus subrufescens fermented rye affects the development of intestinal microbiota, local intestinal and innate immunity in suckling-to-nursery pigs.

BACKGROUND: Agaricus subrufescens is considered as one of the most important culinary-medicinal mushrooms around the world. It has been widely suggested to be used for the development of functional food ingredients to promote human health ascribed to the various properties (e.g., anti-inflammatory, antioxidant, and immunomodulatory activities). In this context, the interest in A. subrufescens based feed ingredients as alternatives for antibiotics has also been fuelled during an era of reduced/banned antibiotics use. This study aimed to investigate the effects of a fermented feed additive -rye overgrown with mycelium (ROM) of A. subrufescens-on pig intestinal microbiota, mucosal gene expression and local and systemic immunity during early life. Piglets received ROM or a tap water placebo (Ctrl) perorally every other day from day 2 after birth until 2 weeks post-weaning. Eight animals per treatment were euthanized and dissected on days 27, 44 and 70. RESULTS: The results showed ROM piglets had a lower inter-individual variation of faecal microbiota composition before weaning and a lower relative abundance of proteobacterial genera in jejunum (Undibacterium and Solobacterium) and caecum (Intestinibacter and Succinivibrionaceae_UCG_001) on day 70, as compared to Ctrl piglets. ROM supplementation also influenced gut mucosal gene expression in both ileum and caecum on day 44. In ileum, ROM pigs showed increased expression of TJP1/ZO1 but decreased expression of CLDN3, CLDN5 and MUC2 than Ctrl pigs. Genes involved in TLR signalling (e.g., TICAM2, IRAK4 and LY96) were more expressed but MYD88 and TOLLIP were less expressed in ROM pigs than Ctrl animals. NOS2 and HIF1A involved in redox signalling were either decreased or increased in ROM pigs, respectively. In caecum, differentially expressed genes between two groups were mainly shown as increased expression (e.g., MUC2, PDGFRB, TOLLIP, TNFAIP3 and MYD88) in ROM pigs. Moreover, ROM animals showed higher NK cell activation in blood and enhanced IL-10 production in ex vivo stimulated MLN cells before weaning. CONCLUSIONS: Collectively, these results suggest that ROM supplementation in early life modulates gut microbiota and (local) immune system development. Consequently, ROM supplementation may contribute to improving health of pigs during the weaning transition period and reducing antibiotics use.

Uptake of Ultrashort Chain, Emerging, and Legacy Per- and Polyfluoroalkyl Substances (PFAS) in Edible Mushrooms (Agaricus spp.) Grown in a Polluted Substrate.

Uptake of 19 per- and polyfluoroalkyl substances (PFAS), including C3-C14 perfluoroalkyl carboxylic acids (PFCAs), C4, C6, and C8 perfluoroalkyl sulfonates (PFSAs), and four emerging PFAS, was investigated in two mushroom species (Agaricus bisporus and Agaricus subrufescens) cultivated in a biogas digestate-based substrate. Accumulation of PFAS in mushrooms was low and strongly chain-length dependent. Among the different PFCAs, bioaccumulation factors (log BAFs) decreased from a maximum of -0.3 for perfluoropropanoic acid (PFPrA; C3) to a minimum of -3.1 for perfluoroheptanoate (PFHpA; C7), with only minor changes from PFHpA to perfluorotridecanoate (PFTriDA; C13). For PFSAs, log BAFs decreased from perfluorobutane sulfonate (PFBS; -2.2) to perfluorooctane sulfonate (PFOS; -3.1) while mushroom uptake was not observed for the alternatives 3H-perfluoro-3-[(3-methoxy-propoxy)propanoic acid] (ADONA) and two chlorinated polyfluoro ether sulfonates. To the best of our knowledge, this is the first investigation of the uptake of emerging and ultra-short chain PFAS in mushrooms, and generally the results indicate very low accumulation of PFAS.

Biochemical Approaches on Commercial Strains of Agaricus subrufescens Growing under Two Environmental Cultivation Conditions.

In the present work, the effect of the cultivation process, in the field and under a controlled environment, on biochemical parameters by using commercial strains of A. subrufescens were evaluated. The results obtained revealed that the strains cultivated in the field presented higher levels for most of the parameters evaluated (organic acids (20.5-48.0 g/100 g dw), tocopherols (107.0-198.6 µg/100 g dw), and phenolic acids and related compounds (245.2-359.0 µg/100 g dw and 10.6-23.7 µg/100 g dw, respectively)), except for the carbohydrates (53.4-72.6 g/100 g dw), energetic value (373-380 Kcal/100 g dw), and total free sugars (28.8-43.1 g/100 g dw), parameters in which the strains grown in a controlled environment present better results. For both cultivation systems, similar results were obtained regarding saturated, monounsaturated, and polyunsaturated fatty acids, as well as antioxidant and antimicrobial activities. These data contribute to the knowledge and highlight the characterized strains and the cultivation process, which can be used to obtain ingredients with potential applicability as a source of functional compounds.

Influence of strains and environmental cultivation conditions on the bioconversion of ergosterol and vitamin D2 in the sun mushroom.

BACKGROUND: The fungus Agaricus subrufescens is grown commercially in China, the USA, Brazil, Taiwan and Japan, among others. However, each country adopts a cultivation system that significantly influences the agronomical parameters and chemical composition of the harvested mushrooms. In this study, the influence of the cultivation process on the content of ergosterol and vitamin D2 was evaluated. RESULTS: Four commercial strains of A. subrufescens (ABL 04/49, ABL CS7, ABL 18/01 and ABL 19/01) and two environmental cultivation conditions (in the field and a controlled chamber with the absence of sunlight) were used. Infield cultivation, ABL CS7 and ABL 19/01 strains presented better agronomic parameters, whereas in a protected environment ABL 19/01, ABL 04/49 and ABL 18/01 demonstrated better performance, respectively. The highest biological efficiency value (64%) was provided by ABL 19/01 strain in a controlled environment. CONCLUSION: The highest content in ergosterol (990 mg kg-1 ) and vitamin D2 (36.8 mg kg-1 ) were observed in mushrooms obtained in the field from strain ABL 04/49, which presents reasonable agronomic parameters for cultivation. © 2021 Society of Chemical Industry.

Changes in layers of laboratory vermicomposting using spent mushroom substrate of Agaricus subrufescens P.

World mushroom production in 2018 was approximately 8.99 million tonnes. The most commonly cultivated species in the Czech Republic are Agaricus spp., which are sold fresh or canned. In 2017, 2018 mushroom production in the Czech Republic was approximately 540 tonnes. Vermicomposting is an easy and less ecologically harmful way to process the spent mushroom substrate. Earthworms, which are referred to as the engine of the process of vermicomposting, and microorganisms, help convert organic waste into fertilizer. This study is concerned with laboratory vermicomposting in a system of continuous feeding of earthworms Eisenia andrei. It compares the differences between variants with and without earthworms. The dry matter percentage was approximately 20% or more in both variants. The variant with earthworms showed a significant decrease in electrical conductivity. The C/N ratio was very low in both variants. The highest total P was recorded in the variant with earthworms, but the highest values of K and Mg were found in the control. Both variants recorded higher content of bacteria than fungi. All values of microorganism contents were higher in the vermicomposter without earthworms, but the bacterial/fungal ratio was higher in the variant with earthworms. The highest content in both variants shows the bacteria especially G-bacteria, on the other side, the lowest content shows the actinobacteria. The highest activity of ß-D-glucosidase and acid phosphatase was measured in the vermicomposter with earthworms, but the activity of other enzymes was higher in the control. In both vermicomposters laccase activity was below the detection limit. The method of classical vermicomposting can be used for processing the spent mushroom substrate. However, in terms of higher content of total and available nutrients, there seems to be a better method of processing the substrate without earthworms.

Cadmium mobility in a circular food-to-waste-to-food system and the use of a cultivated mushroom (Agaricus subrufescens) as a remediation agent.

Cadmium (Cd) and its mobility and toxicity in the food chain receives considerable public and scientific attention. Levels in soil have been increasing, partly caused by mineral fertiliser use. In many areas in the EU, intake of Cd is not far from the maximum tolerable level. Now, urban and more circular approaches to food production are being promoted, in which waste materials substitute growing media and fertilizers. Can this make the situation worse, or is it, on the contrary, possible to extract Cd from the cycle using practical microbial methods? Cd concentration, availability and accumulation in materials and crops in a closed greenhouse system was investigated first. Secondly, we tested if fungal remediation was able to remove a significant amount of Cd. A sequential extraction analysis showed how anaerobic digestion and composting changed chemical Cd availability. Cd levels in the system were comparable to agricultural soils. The vegetable crops did not accumulate Cd. However, the culture of the edible mushroom Agaricus subrufescens in the greenhouse had a strong effect on Cd mobility: It extracted 80% of the Cd from the growing medium into the mushrooms, mostly at the start of fruiting. These early Cd enriched mushrooms can be treated as special waste, while the later harvest complied with EU Cd limits for foodstuffs and can potentially be consumed. Thus, growing this mushroom species in an integrated waste-to-crop system can be a cost-efficient way to remediate Cd from the food chain.

Cereal grains and glycerol in Agaricus blazei cryopreservation / Grãos de cereais e glicerol na criopreservação de Agaricus blazei

Agaricus blazei is an important commercial basidiomycete studied for its biological activity. However, little has been studied about the preservation techniques of this basidiomycete, and cryopreservation is one alternative. The aim of this study was to evaluate the physical and chemical effects of different cultivation media and glycerol on A. blazei cryopreservation at -20 ºC and at -70 ºC. A solid cultivation medium consisting of agar with ground whole cereal grains (hard endosperm wheat, medium hard endosperm wheat or hard endosperm rye) or malt extract agar, or whole cereal grains (hard endosperm wheat, medium hard endosperm wheat or hard endosperm rye) without agar was used. Cultivation media had zero or 5% glycerol addition. Cultivation medium grains or disks containing mycelia were transferred to cryotubes with glycerol at 5% with or without water and then were cryopreserved at -20 ºC or at -70 ºC. After one-year or two-year cryopreservation, fungus viability was evaluated. The results showed that the physical structure of whole grains of hard and medium hard wheat is effective (p≤0.01) for two-year cryopreservation at -70 ºC as well as the use of glycerol in the cultivation medium or as a cryoprotectant (p≤0.01) at -70 ºC. Cryopreservation at -20 ºC was ineffective (p≤0.01) to preserve fungus viability.

Agaricus blazei é um basidiomiceto de importância comercial amplamente estudado quanto à sua atividade biológica. Entretanto, pouco foi estudado sobre as técnicas de preservação deste basidiomiceto, sendo a criopreservação uma alternativa. O objetivo deste trabalho foi avaliar os efeitos físicos e químicos de diferentes meios de cultivo e do glicerol na criopreservação de A. blazei à -20 ºC e à -70 ºC. Foi usado meio de cultivo sólido composto por ágar com farinha de grãos inteiros de cereais (trigo textura dura ou semidura ou centeio textura dura) ou ágar extrato de malte, ou grãos de cereais inteiros (trigo textura dura ou semidura ou centeio textura dura) sem ágar. Os meios de cultivo tiveram adição de glicerol (zero ou 5%). Grãos ou discos de meio de cultivo contendo o micélio foram transferidos para criotubos com glicerol a 5%, ou com ou sem água, sendo, em seguida, criopreservados (-20 ºC ou -70 ºC). Após um ou dois anos de criopreservação foi avaliada a viabilidade do fungo. Os resultados mostraram que a estrutura física de grãos inteiros de trigo semiduro ou duro é efetiva (p≤0,01), quando em -70 ºC para criopreservação por dois anos, assim como o uso de glicerol tanto no meio de cultivo ou como solução crioprotetora (p≤0,01). A criopreservação a -20 ºC mostrou-se ineficaz (p≤0,01) na manutenção da viabilidade do fungo.

Cryopreservation of Agaricus blazei in liquid nitrogen using DMSO as cryoprotectant / Criopreservação de Agaricus blazei em nitrogênio líquido usando DMSO como crioprotetor

The preservation of Agaricus blazei is generally done using successive subcultivations that are laborious and are subject to contaminations or genetic degenerations, resulting in loss of biotechnological interest characteristics. An alternative process would be cryopreservation, but there are no reports of methodologies for this basidiomycete in liquid nitrogen. Thus, the objective of this study was to evaluate mycelial viability of A. blazei strains after cryopreservation in liquid nitrogen in order to establish the initial parameters of species preservation. Five strains grown on malt extract agar (MEA) were used. Disks of MEA containing A. blazei mycelium were transferred for screwcap cryovials containing the cryoprotectant, 10% dimethyl sulfoxide. Then, they were cooled at 8 ºC for 30 min and kept at -196 ºC with liquid nitrogen. After 1.5 year of cryopreservation, the cryovials were thawed in water bath at 30 ºC for 15 min. The disks with mycelia were transferred to MEA culture media without cryoprotectant and kept at 28 ºC for 30 days. A. blazei strains respond differently to the cryopreservation method at -196 ºC by varying mycelial viability recovery. Cryopreservation with liquid nitrogen, using dimethyl sulfoxide as cryoprotectant, is not the most appropriate one for A. blazei preservation.

A preservação de Agaricus blazei ocorre usualmente por subculturas sucessivas que são laboriosas e estão sujeitas a contaminações ou degenerações genéticas, com eventual perda das características de interesse biotecnológico. Uma alternativa a este processo é a criopreservação, no entanto não existem relatos de metodologias para este basidiomiceto com uso de nitrogênio líquido. Desta forma, o objetivo deste trabalho foi avaliar a criopreservação em nitrogênio líquido de linhagens de A. blazei, visando estabelecer os parâmetros iniciais de criopreservação para a espécie. Foram utilizadas cinco linhagens do fungo crescidas em meio de ágar-extrato de malte (MEA). Os discos de MEA contendo o micélio crescido foram transferidos em temperatura ambiente (25 ºC) para criotubos com rosca contendo o agente crioprotetor dimetilsulfóxido a 10%. Em seguida foram resfriados a 8 ºC por 30 min e mantidos a -196 ºC com nitrogênio líquido. Após 1,5 anos de criopreservação os criotubos foram descongelados por imersão em água a 30 ºC por 15 min. Os discos com micélio foram transferidos para meio de cultura MEA, sem o agente crioprotetor, e mantidos por 30 dias a 28 ºC. As linhagens de A. blazei respondem de forma diferente ao processo de criopreservação a -196 ºC com variação na recuperação da viabilidade do micélio. A criopreservação em nitrogênio líquido de A. blazei, com dimetilsulfóxido como crioprotetor, não é a forma mais adequada para a preservação desta espécie.

Production flush of Agaricus blazei on Brazilian casing layers

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

Production flush of Agaricus blazei on Brazilian casing layers.

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