Effect of Agaricus blazei Murill on exploratory behavior of mice-model / Efeito de Agaricus blazei Murill no comportamento exploratório de modelo de camundongo

Abstract Increased anxiety and depressive symptoms have reported to be its association with long term illness. Because of having unwanted effects of newly available drugs, patients administering anxiolytic drugs usually discontinue the treatment before they are completely recovered. Therefore, there is a serious need to develop new anxiolytic drugs. The anxiolytic effect of hydro-alcoholic extract of Agaricus blazei in animal models was assessed. 24 male mice (Mus musculus genus) were included in the study. Four groups were prepared and each group contained six animals. The groups were vehicle control, positive control (diazepam 1.0 mg/kg, i.p.) as well as two treatment groups receiving Agaricus blazei hydro-alcoholic extract at a dose of 136.50 mg/kg and 273.0 mg/kg orally. The Marble burying test, Nestlet shredding test and Light and Dark box test used to assess anxiolytic activity. Mice administered with diazepam 1.0 mg/kg, i.p. while hydro-alcoholic extract of AbM (136.50 and 273.0 mg/kg, respectively) was administered via oral route which exhibited marked reduction in number of marbles-burying as compared to vehicle control group. Mice administered with diazepam 1.0 mg/kg, i.p. and Oral administration of hydro-alcoholic extract of AbM (136.50 and 273.0 mg/kg, respectively) exhibited significant decrease in nestlet shredding in comparison to vehicle control group. The oral administration of hydro-alcoholic extract at a dose of 136.5mg/kg and 273mg/kg showed elevation in time spent in light box and was comparable to standard treated group while time spent by mice following oral administration of hydro-alcoholic extract of Agaricus blazei at a dose of 273.0 mg/kg also showed elevation and was found to be more near to standard treated group (diazepam 1 mg/kg, i.p.).

Resumo O aumento da ansiedade e dos sintomas depressivos têm relatado sua associação com doenças de longa duração. Por causa dos efeitos indesejáveis dos novos medicamentos disponíveis, os pacientes que administram medicamentos ansiolíticos geralmente interrompem o tratamento antes de estarem completamente recuperados. Portanto, há uma necessidade séria de desenvolver novos medicamentos ansiolíticos. Foi avaliado o efeito ansiolítico do extrato hidroalcoólico de Agaricus blazei em modelos animais. Vinte e quatro camundongos machos (gênero Mus musculus) foram incluídos no estudo. Quatro grupos foram preparados, e cada grupo continha seis animais. Os grupos foram controle de veículo, controle positivo (diazepam 1,0 mg/kg, i.p.), bem como dois grupos de tratamento recebendo extrato hidroalcoólico de Agaricus blazei na dose de 136,50 mg/kg e 273,0 mg/kg por via oral. O teste de enterrar Marble, o teste de retalhamento Nestlet e o teste de caixa clara e escura são usados ​​para avaliar a atividade ansiolítica. Camundongos foram administrados com diazepam 1,0 mg/kg, i.p., enquanto o extrato hidroalcoólico de AbM (136,50 e 273,0 mg/kg, respectivamente) foi administrado por via oral, que exibiu redução acentuada no número de mármores enterrados em comparação com o grupo de controle de veículo. Camundongos administrados com diazepam 1,0 mg/kg, i.p. e a administração oral de extrato hidroalcoólico de AbM (136,50 e 273,0 mg/kg, respectivamente) exibiu diminuição significativa na trituração de ninhos em comparação ao grupo de controle de veículo. A administração oral de extrato hidroalcoólico na dose de 136,5mg/kg e 273mg/kg mostrou elevação no tempo gasto na caixa de luz e foi comparável ao grupo tratado padrão, enquanto o tempo gasto por camundongos após a administração oral de extrato hidroalcoólico de Agaricus blazei na dose de 273,0 mg/kg também mostrou elevação e foi mais próximo do grupo tratado padrão (diazepam 1 mg/kg, ip).

Variability in growth and biofilm formation of Listeria monocytogenes in Agaricus bisporus mushroom products.

Foods and food production environments can be contaminated with Listeria monocytogenes and may support growth of this foodborne pathogen. This study aims to characterize the growth and biofilm formation of sixteen L. monocytogenes strains, isolated from mushroom production and processing environments, in filter-sterilized mushroom medium. Strain performance was compared to twelve L. monocytogenes strains isolated from other sources including food and human isolates. All twenty-eight L. monocytogenes strains showed rather similar growth performance at 20 °C in mushroom medium, and also significant biofilm formation was observed for all strains. HPLC analysis revealed the presence of mannitol, trehalose, glucose, fructose and glycerol, that were all metabolized by L. monocytogenes, except mannitol, in line with the inability of L. monocytogenes to metabolize this carbohydrate. Additionally, the growing behavior of L. monocytogenes was tested on whole, sliced and smashed mushroom products to quantify performance in the presence of product-associated microbiota. A significant increase of L. monocytogenes was observed with higher increase of counts when the mushroom products were more damaged, even with the presence of high background microbiota counts. This study demonstrated that L. monocytogenes grows well in mushroom products, even when the background microbiota is high, highlighting the importance to control (re)contamination of mushrooms.

Insights into the genomic evolution and the alkali tolerance mechanisms of Agaricus sinodeliciosus by comparative genomic and transcriptomic analyses.

Agaricus sinodeliciosus is a rare wild edible mushroom from northwest China, and grows naturally in mild saline-alkali soil, which is also unusual in mushrooms. A. sinodeliciosus represents a potential model organism for explaining saline-alkali tolerance mechanisms and revealing related physiological processes in mushrooms. Here, we provide a high-quality genome of A. sinodeliciosus. Comparative genomic analyses reveal A. sinodeliciosus has numerous changes to its genome organization after a solitary evolutionary history under saline-alkali environments, such as gene family contraction, retrotransposon expansion and rapid evolution of adaptative genes. Our saline and alkali tolerance tests show that mycelium growth and fruit body formation of this species are effected by mild alkalinity. Transcriptomic analyses reveal that genes involved in carbon and nitrogen utilization, cell stability and fruit body formation of A. sinodeliciosus could be activated under mildly alkaline conditions. In particular, the 'starch and sucrose metabolism', 'biosynthesis of amino acids' and 'phenylpropanoid biosynthesis' pathways are important for mildly alkaline tolerance of A. sinodeliciosus. Like plants and arbuscular mycorrhizal fungi, in the rot fungus A. sinodeliciosus, the biosynthesis of intracellular small molecules could be enhanced to counter osmotic and oxidative stresses caused by mild alkalinity, and the biosynthesis of monolignol could be suppressed to increase cell wall infiltrates under mildly alkaline conditions. This research provides an understanding of the genomic evolution and mechanisms of A. sinodeliciosus in tolerance to saline-alkali environments. The A. sinodeliciosus genome constitutes a valuable resource for evolutionary and ecological studies of Agaricus.

Agaricus bisporus Wild Mushroom Extract as Lectin Source for Engineering a Lactose Photoelectrochemical Biosensor.

Agaricus bisporus mushroom biomass contains a lectin, ABL, with remarkable specificity for lactose biorecognition; in this work, this feature was explored to develop a photoelectrochemical biosensor. The high lectin activity found in saline extracts of this macrofungus (640 HU mL-1), even at critical pH values (4-10) and temperatures (20-100 °C), allowed its direct use as an ABL source. Theoretical and experimental evidence revealed favorable electrostatic and biocompatible conditions to immobilize ABL on a poly(methylene blue)/fluorine-doped tin oxide-coated glass platform, giving rise to the ABL/PMB/FTO biosensor. The conducting polymer added further photoactivity to the device, allowing the identification of lectin-carbohydrate interactions with even greater sensitivity. The dose-response curves studied by electrochemical impedance spectroscopy showed a sigmoidal profile that was well-fitted by Hill's equation, expanding the working dynamic range (15-540 nmol L-1 lactose; 20.2 pmol L-1 detection limit) and avoiding undesirable sample dilution or preconcentration procedures. Under the optimized photoelectrochemical conditions, the ABL/PMB/FTO biosensor showed remarkable signal stability, accuracy, specificity, and selectivity to analyze lactose in commercial food products. This research raises interest in ABL-based biosensors and the added value of the crude Agaricus bisporus extract toward the development of greener and more sustainable biotechnological approaches.

Isolation, Characterization, and Biocompatibility of Bisporitin, a Ribotoxin-like Protein from White Button Mushroom (Agaricus bisporus).

White button mushroom (Agaricus bisporus (J.E. Lange) Imbach) is one of the widely consumed edible mushrooms. Indeed, A. bisporus fruiting bodies are a rich source of nutrients and bioactive molecules. In addition, several enzymes with biotechnological applications are found in A. bisporus (e.g., enzymes for lignocellulose degradation). Here, a novel ribotoxin-like protein (RL-P) from the edible mushroom A. bisporus was purified and characterized. This RL-P, named bisporitin, is a monomeric protein (17-kDa) exhibiting specific ribonucleolytic activity by releasing the α-fragment (hallmark of RL-Ps) when incubated with rabbit ribosomes. In addition, bisporitin shows magnesium-dependent endonuclease activity and displays a similar far-UV CD spectrum as ageritin, the prototype of RL-Ps, isolated from Cyclocybe aegerita fruiting bodies. Interestingly, bisporitin is the first member of RL-Ps to have noticeably lower thermal stability (Tm = 48.59 ± 0.98 °C) compared to RL-Ps isolated in other mushrooms (Tm > 70 °C). Finally, this protein is only partially hydrolyzed in an in vitro digestive system and does not produce adverse growing effects on eukaryotic cell lines. This evidence paves the way for future investigations on possible bioactivities of this RL-P in the digestive system.

Influence of Agaricus bisporus Mushroom on Pb Toxicokinetic in Pregnant Rats.

(Pb) is a toxic metal, responsible for several damages to human health. Agaricus bisporus (Ab) is a mushroom with promising antioxidant properties to be used as an alternative chelator in Pb intoxication. The aim was to understand the Pb toxicokinetic and the potential of Ab as a protective agent. A total of 20 female Wistar rats were distributed into 4 groups (n = 5/group): Control (receiving water); Group Ab 100 mg/kg (gavage); Group Pb 100 mg/L in water; and Group Ab + Pb-100 mg/kg + 100 mg/L (gavage and water). Pb administration occurred daily until the 19th day of pregnancy. On day 19 of gestation, the rats were euthanized, and the blood and tissues were collected for Pb measurement, using an inductively coupled plasma mass spectrometer. The results showed that the levels of Pb in the blood, placenta, and liver of the mothers, and in the brain of the fetuses increased significantly in the Pb group. On the other hand, the combined exposure to Pb + Ab showed a significant decrease in the metal concentration in relation to the Pb group, returning to normal levels. Kidney and bone lead levels also increased significantly in the Pb group. However, in the combined exposure group, levels did not return to the control amounts; there was protection, but the Pb concentration was still significantly higher than in the control. In the brain, no significant differences were observed. In conclusion, we suggest A. bisporus is a natural chelator, because the co-administration of the mushroom was able to interact with Pb ions, minimizing the Pb absorption and distribution. These effects are suggested since A. bisporus have antioxidants and beta glucan that interact with Pb, chelating it and, thus, reducing its toxic effects.

Distribution of Virulence Genes in Campylobacter spp. Isolated from Agaricus Mushrooms in Iran.

The white button mushroom (Agaricus) is a significant nutritional and therapeutic species utilized in the human diet and could transmit various bacterial infections. Campylobacter species are the most common cause of foodborne illness across the world. The present study has been planned to determine the frequency of virulence genes and antibiotic susceptibility test in Campylobacter spp. recovered from Agaricus mushroom. In this study, 740 Agaricus mushroom samples were gathered randomly from various markets from June 2020 to December 2020. Confirmation of Campylobacter spp. using biochemical analyses and 23S rRNA-based PCR was performed. The agar dilution technique was used to determine resistance to antibiotics using gentamicin (GM10µg), ciprofloxacin (CIP5µg), nalidixic acid (NA30µg), tetracycline (TE30µg), ampicillin (AM10µg), amoxicillin+ clavulanic acid (AMC30µg), erythromycine (E15µg), azithromycin (AZM15µg), clindamycin (CC2µg), and chloramphenicol (C30µg). Multiplex PCR was utilized to determine the prevalence of the recR, dnaJ, wlaN, virBll, cdtC, cdtB, cdtA, flaA, cadF, pidA, ciaB, ceuE, and cgtB genes. Campylobacter spp. were detected in 74 out of 740 Agaricus mushroom samples (10%). According to the data, Agaricus mushroom samples included 32 (4.32%) C. jejuni, 11 (1.48%) C. coli, and 31 (4.18%) other Campylobacter spp. Antimicrobial resistance was most common in C. jejuni isolates. C. jejuni isolates also had the lowest resistance rate to gentamycin, ciprofloxacin, and nalidixic acid. C. coli isolates were reported to have the highest antimicrobial resistance to ciprofloxacin, ampicillin, and erythromycine. Resistance to gentamycin and amoxicillin+ clavulanic acid was likewise lowest among C. coli strains. The flaA and ciaB genes were found in 100% of B-lactams-susceptible C. jejuni and C. coli strains. When examining the relationship between antibiotic resistance and the existence of virulence genes, it was observed that there is a statistically significant relationship (p < 0.001) between bacterial resistance and virulence genes. Our findings indicated that changes in resistance patterns in Campylobacter strains have emerged from multiple treatment approaches in Agaricus mushrooms.

Bacterial interactions with the mycelium of the cultivated edible mushrooms Agaricus bisporus and Pleurotus ostreatus.

The cultivated edible mushrooms Agaricus bisporus and Pleurotus ostreatus are valuable food crops and an important source of human nutrition. Agaricus bisporus is the dominant cultivated species in the western hemisphere and in Australia, while in Asian countries P. ostreatus is more prevalent. These two mushroom species are grown on fermented-pasteurized substrates, and bacteria and fungi play an important role in converting feedstocks into a selective medium for the mushroom mycelium. The mushrooms are usually introduced to the substrate as grain spawn, and the actively growing hyphae form a range of direct interactions with the diverse bacterial community in the substrate. Of these interactions, the most well studied is the removal of inhibitory volatile C8 compounds and ethylene by pseudomonads, which promotes mycelium growth and stimulates primordia formation of both A. bisporus and P. ostreatus. Bacterial biomass in the substrate is a significant nutrition source for the A. bisporus mycelium, both directly through bacteriolytic enzymes produced by A. bisporus, and indirectly through the action of extracellular bacterial enzymes, but this is less well studied for P. ostreatus. Apart from their role as a food source for the growing mycelium, bacteria also form extensive interactions with the mycelium of A. bisporus and P. ostreatus, by means other than those of the removal of inhibitory compounds. Although several of these interactions have been observed to promote mycelial growth, the proposed mechanisms of growth promotion by specific bacterial strains remain largely uncertain, and at times conflicting. Bacterial interactions also elicit varying growth-inhibitory responses from A. bisporus and P. ostreatus. This review explores characterized interactions involving bacteria and A. bisporus, and to a lesser degree P.ostreatus, and whilst doing so identifies existing research gaps and emphasizes directions for future work.

Inhibition of Tolaasin Cytotoxicity Causing Brown Blotch Disease in Cultivated Mushrooms Using Tolaasin Inhibitory Factors.

Tolaasin, a pore-forming bacterial peptide toxin secreted by Pseudomonas tolaasii, causes brown blotch disease in cultivated mushrooms by forming membrane pores and collapsing the membrane structures. Tolaasin is a lipodepsipeptide, MW 1985, and pore formation by tolaasin molecules is accomplished by hydrophobic interactions and multimerizations. Compounds that inhibit tolaasin toxicity have been isolated from various food additives. Food detergents, sucrose esters of fatty acids, and polyglycerol esters of fatty acids can effectively inhibit tolaasin cytotoxicity. These chemicals, named tolaasin-inhibitory factors (TIF), were effective at concentrations ranging from 10-4 to 10-5 M. The most effective compound, TIF 16, inhibited tolaasin-induced hemolysis independent of temperature and pH, while tolaasin toxicity increased at higher temperatures. When TIF 16 was added to tolaasin-pretreated erythrocytes, the cytotoxic activity of tolaasin immediately stopped, and no further hemolysis was observed. In the artificial lipid bilayer, the single-channel activity of the tolaasin channel was completely and irreversibly blocked by TIF 16. When TIF 16 was sprayed onto pathogen-treated oyster mushrooms growing on the shelves of cultivation houses, the development of disease was completely suppressed, and normal growth of oyster mushrooms was observed. Furthermore, the treatment with TIF 16 did not show any adverse effect on the growth of oyster mushrooms. These results indicate that TIF 16 is a good candidate for the biochemical control of brown blotch disease.

Changes in Arsenic Speciation in Wild Edible Fungi after Different Cooking Processes and Gastrointestinal Digestion.

Arsenic (As) is enriched in wild edible fungi, which is one of the main important sources of As in humans' diet. In this study, two wild edible fungi were employed for investigation: (1) Pleurotus citrinopileatusone, which contains a high content of inorganic As (iAs) and (2) Agaricus blazei Murill, which contains a high content of organic As. This study investigated the changes in As content and its speciation after different daily cooking methods. We found that the content of As in Pleurotus citrinipileatus and Agaricus blazei Murill reduced by soaking plus stir-frying by 55.4% and 72.9%, respectively. The As content in Pleurotus citrinipileatus and Agaricus blazei Murill decreased by 79.4% and 93.4%, respectively, after soaking plus boiling. The content of As speciation in dried wild edible fungi reduced significantly after different treatments. Among them, iAs decreased by 31.9~88.3%, and organic As decreased by 33.3~95.3%. This study also investigated the bioaccessibility of As in edible fungi after different cooking processes via an in-vitro physiologically based extraction test (PBET). The results showed that the bioaccessibility of As was relatively high if the edible fungi were uncooked, boiled, or stir-fried. The gastric (G) bioaccessibility of As ranged from 51.7% to 93.0% and the gastrointestinal (GI) bioaccessibility of As ranged from 63.5% to 98.1%. Meanwhile, the bioaccessibility of inorganic As was found to be as high as 94.6% to 151%, which indicates that further evaluation of the potential health risks of wild edible fungi is necessary.

Identification of bHLH family genes in Agaricus bisporus and transcriptional regulation of arginine catabolism-related genes by AbbHLH1 after harvest.

Basic helix-loop-helix (bHLH) transcription factors (TFs) are widely distributed in eukaryotes and play an important role in biological growth and development. The identification and functional analyses of bHLH genes/proteins in edible mushrooms (Agaricus bisporus) have yet to be reported. In the present study, we identified 10 putative bHLH members carrying the conserved bHLH domains. Phylogenetic analyses revealed that the 10 AbbHLHs were the closest to sequences of species belonging to 7 different fungal subgroups, which was supported by loop length, intron patterns, and key amino acid residues. The substantial increase after harvest and continuously elevated expression of AbbHLH1 during the development until the disruption of mushroom velum, and the preferential expression in cap and gill tissues suggest the important function of AbbHLH1 in postharvest development of A. bisporus. The relationship of arginine catabolism-related genes with the early stage of postharvest continuing development also was revealed by expression determination. Subcellular localization showed that AbbHLH1 could be localized in nucleus. Importantly, the electrophoretic mobility shift and dual-luciferase reporter assays showed that AbbHLH1 activated the promoters of AbOAT, AbSPDS, and AbSAMDC and suppressed the expression of AbARG, AbUREA, and AbODC, probably for the modulation of arginine catabolism and thus control of postharvest mushroom development. Taken together, the available data provide valuable functional insight into the role of AbbHLH proteins in postharvest mushrooms.

Bacterial microbiota profiling of oyster mushrooms (Pleurotus ostreatus) based on cultivation methods and distribution channels using high-throughput sequencing.

The annual consumption and production of oyster mushrooms (Pleurotus ostreatus) have continued to rise due to its nutritive and health-promoting benefits. Cultivated mushrooms are mostly grown in small to medium-scaled scale production plants that present hygienic challenges which could, in turn, increase associated foodborne pathogenic outbreaks. The present study aimed to investigate the shift in microbial ecologies of oyster mushrooms from pre-distribution (cultivation in bottles or on shelves) to post-distribution at supermarkets and open-air markets. Aerobic plate counts and coliforms were quantified using traditional microbiological techniques, and the microbiome associated with oyster mushrooms (n = 70) was analyzed using 16S rRNA amplicon sequencing for an enhanced level of bacterial microbiota profiling. Overall, coliforms recovered from pre-distribution bottle-cultivated mushrooms were 1.9 log CFU/g higher (p < 0.05) than that of shelf-cultivated mushrooms. The mean aerobic plate counts of oyster mushrooms distributed to open-air markets was 1.2 log CFU/g higher (p < 0.05) than packaged mushrooms from supermarkets while there were no significant differences in coliform counts. The pattern of bacterial composition differed by post-distribution channels, with oyster mushrooms collected from the open-air markets demonstrating the richest microbiome diversity. An increase in the relative abundance of Enterobacteriaceae (55-68 %) and Pseudomonadaceae (27-35 %) was observed in pre- and post-distribution mushrooms, respectively. However, no distinct bacterial microbiota differences were observed for the different cultivation methods or different geographical locations for each market type. The current findings add to our understanding of the effects of cultivation methods and commercial distribution channels regarding the microbiome of oyster mushrooms and may inform potential intervention strategies for future production and distribution processes. Furthermore, the tandem analyses of culture-dependent and culture-independent methods can provide more comprehensive information than that obtained when using each approach independently.

Anti-phytopathogenic-bacterial fatty acids from the mycelia of the edible mushroom Agaricus blazei.

Five compounds including a new compound (1) were isolated from mycelia of a mushroom-forming fungus Agaricus blazei. Compound 2 was isolated from nature for the first time. Their structures were determined by the interpretation of spectroscopic data. In the bioassay examining growth inhibitory activity against phytopathogenic bacteria Clavibacter michiganensis, Burkholderia glumae, and Peptobacterium carotovorum, all the compounds showed inhibition effects on C. michiganensis. Compounds 3 and 4 also showed weak inhibitory activity against growth of B. glumae.

Modulation of obesity associated metabolic dysfunction by novel lipophilic fraction obtained from Agaricus bisporus.

AIM: The present study aimed to prepare a novel lipophilic fraction rich in fat soluble bioactive from Agaricus bisporus and investigated its impact through in vitro and in vivo assessments since the prospective biological activities of fat soluble components from mushrooms are limited. METHODS: Ergosterol concentrate fraction (ECF) preparation followed by subsequent characterization of the extract using various analytical techniques (HPLC-UV, Mass spectroscopy, NMR). Furthermore, the fraction has been evaluated for antioxidant activity, DNA protection ability, hypolipidemic properties by in vitro specific enzyme inhibition and in vivo animal model (C57BL/6). KEY FINDINGS: The fraction majorly contains ergosterol (504 mg/100 g dw) and linoleic acid (71.92 %). In vitro studies showed that the fraction limited free radicals induced DNA damage, exhibited significant free radical scavenging activities (IC50 of DPPH 15.64; ABTS 8.28 mg/ml), and inhibited HMG-CoA reductase activity (IC50 5.03 mg/ml). Further, in vivo study showed that ECF treatment significantly (p < 0.05) improved insulin sensitivity (reduced plasma glucose & insulin, increased adiponectin) and reduced inflammatory markers (CRP & TNF-α) in comparison to high fat fed mice. Furthermore, ECF has significantly reduced plasma lipid profile and accumulation of lipids in liver. This could be due to down regulation of mRNA expression of lipogenic transcription factors such as SREBP-1c and SREBP-2, and key lipogenic enzyme ACC. Moreover, ECF treatment has suppressed protein expression of FAS, induced cholesterol clearance by enhancing LDL-R protein expression. SIGNIFICANCE: The present work for the first time evaluated the synergistic potential of ergosterol and linoleic acid to improve antioxidant defense system and ameliorate obesity associated metabolic dysfunction.

Immunomodulatory activity of extracts from five edible basidiomycetes mushrooms in Wistar albino rats.

Mushrooms are nutritious foods that are widely cultivated all over the world. They are rich in a range of compounds linked to improving functions of the immune system including carotenoids, alkaloids, lectins, enzymes, folates, fats, organic acids, minerals, polysaccharides, phenolics, proteins, tocopherols, terpenoids, and volatile compounds. In this study we investigated, the immunomodulatory activity in rats of the aqueous extracts of five of the most common edible mushrooms belonging to Family Basidiomycota-white-rot fungi including, Lentinula edodes, Agaricus bisporus, Pleurotus ostreatus, Pleurotus columbinus, and Pleurotus sajor-caju. Male Wistar albino rats were assigned to thirteen groups and Immunosuppression was induced by oral administration of dexamethasone (0.1 mg/kg), followed by oral administration of the mushroom extracts at low (200 mg/kg) and high (400 mg/kg) doses. A positive control group received the immune stimulant Echinacea extract Immulant® at (30 mg/kg), while the negative control group received only saline. From each animal, in each group, blood samples were collected after 15 days for complete blood counts and for measurement of immunologic parameters, including lysozyme activity, nitric oxide (NO) production and serum cytokines including tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ) and interleukin 1 beta (IL-1ß) levels. Results have shown that white blood cells (WBCs) and lymphocytic counts were significantly boosted by high doses of each of the five mushroom extracts (207-289% increase for WBC and 153-175% for lymphocytes) with a significant increase in lysozyme activity (110-136% increase), NO concentration (159-232% increase) and cytokines as compared to the negative control group. Histopathological examination of the rats' spleen and thymus tissues has shown marked lymphocytic proliferation that was more obvious at the higher doses. In conclusion, our results showed that the five edible mushroom extracts revealed significant immunostimulatory effects preclinically particularly, at the higher doses (400 mg/kg) which can be considered the effective dose.

Caffeic acid-grafted-chitosan/polylactic acid film packaging enhances the postharvest quality of Agaricus bisporus by regulating membrane lipid metabolism.

In this study, the film assembled by caffeic acid-grafted-chitosan/polylactic acid (CA-g-CS/PLA) was used for packaging postharvest Agaricus bisporus, and its effects on postharvest quality of Agaricus bisporus by regulating membrane lipid metabolism were investigated. The results showed that compared to traditional polyethylene packaging, the cell morphology of Agaricus bisporus packaged by CA-g-CS/PLA demonstrated a more complete structure due to its lower permeability, higher fluidity and stronger ability to maintain free water. Meanwhile, CA-g-CS/PLA packaging could effectively delay the decrease rate of unsaturation value and phospholipids of Agaricus bisporus during storage. In addition, the activities and expression levels of lipoxygenase (LOX), phospholipase C (PLC) and phospholipase D (PLD) in Agaricus bisporus packaged with CA-g-CS/PLA were inhibited, while those of glycerol-3-phosphate acyltransferase (GPAT) and sphingomyelin synthase (SMS) were promoted. Therefore, CA-g-CS/PLA packaging could enhance the postharvest quality of Agaricus bisporus by regulating membrane lipid metabolism, which has great application potential in the field of new intelligent packaging.

Feeding growing button mushrooms: The role of substrate mycelium to feed the first two flushes.

A number of experiments were done to further our understanding of the substrate utilization in button mushroom crops (Agaricus bisporus). An analysis of the degradation of dry matter of the substrate during a crop cycle revealed that for pin formation the upper 1/3rd layer is used, for the production of flush one all layers are involved and for flush two mainly the lower 1/3 layer is used. A reduction in substrate depth leads to a decrease in yield/m2 but an apparent increase in yield per tonne of substrate with a lower mushroom quality. A short daily interruption of the connection between the casing soil with the substrate results in a delay of the first flush. Interruptions with only part of the substrate did not lead to delay in production. Daily interruption of the connection with all or only part of the substrate leads to a shift in yield from flush one to flush two but the total yield remains unchanged. The mycelial biomass in the substrate increases from filling up to pinning, has a steeper increase during flush one, and is levelling off during flush two, indicating that in the period of venting and up to/including flush one, enzymes are secreted by growing hyphae generating nutrients to feed a fixed amount of mushroom biomass for two flushes. A sidewise extension of the substrate (without casing soil, thus not producing mushrooms) showed that the substrate at a distance more than somewhere between 20-50 cm away from the casing soil does not contribute to feeding mushrooms in the first two flushes. The observations are discussed with respect to relevant previous research.

Response of the mushroom pathogen Cladobotryum mycophilum to prochloraz and metrafenone fungicides and Streptomyces flavovirens actinobacteria.

After an outbreak of cobweb disease of cultivated button mushroom in Serbia in 2003, the isolated fungal pathogen was initially identified as Cladobotryum dendroides (teleomorph Hypomyces rosellus) based on morpho-physiological traits. Molecular analysis indicated re-classification of two strains (isolated in 2004 and 2007) as Cladobotryum mycophilum (teleomorph Hypomyces odoratus). However, subsequent analysis of further five strains (isolated over the period 2003-2010) within the frames of the present study, also confirmed their identification as the exclusive cobweb causal agent C. mycophilum. After artificial inoculation, the symptoms observed on harvested and growing mushrooms were consistent with the appearance of cobweb disease. Pathogen sensitivity to fungicides was estimated by probit analyses. Fungicide susceptibility tests showed that C. mycophilum strains were highly sensitive both to prochloraz (ED50<0.087 µg mL-1) and the newly introduced metrafenone (ED50<0.15 µg mL-1). Furthermore, the growth of all examined strains of C. mycophilum was significantly inhibited by the indigenous actinobacterial strain Streptomyces flavovirens A06. A dual culture assay showed after 72 h that the percentage of radial growth inhibition of the pathogen ranged from 22.38 to 55.73%. Our findings suggest that the antagonistic S. flavovirens A06 might be a potential candidate for controlling the cobweb disease of cultivated button mushroom.

Influence of Agaricus bisporus establishment and fungicidal treatments on casing soil metataxonomy during mushroom cultivation.

The cultivation of edible mushroom is an emerging sector with a potential yet to be discovered. Unlike plants, it is a less developed agriculture where many studies are lacking to optimize the cultivation. In this work we have employed high-throughput techniques by next generation sequencing to screen the microbial structure of casing soil employed in mushroom cultivation (Agaricus bisporus) while sequencing V3-V4 of the 16S rRNA gene for bacteria and the ITS2 region of rRNA for. In addition, the microbiota dynamics and evolution (bacterial and fungal communities) in peat-based casing along the process of incubation of A. bisporus have been studied, while comparing the effect of fungicide treatment (chlorothalonil and metrafenone). Statistically significant changes in populations of bacteria and fungi were observed. Microbial composition differed significantly based on incubation day, changing radically from the original communities in the raw material to a specific microbial composition driven by the A. bisporus mycelium growth. Chlorothalonil treatment seems to delay casing colonization by A. bisporus. Proteobacteria and Bacteroidota appeared as the most dominant bacterial phyla. We observed a great change in the structure of the bacteria populations between day 0 and the following days. Fungi populations changed more gradually, with A. bisporus displacing the rest of the species as the cultivation cycle progresses. A better understanding of the microbial communities in the casing will hopefully allow us to increase the biological efficiency of the crop.

The cultural role played by the ethnomycological knowledge of wild mushrooms for the peoples of highlands and lowlands in Tlaltenango, Zacatecas, Mexico.

The greatest diversity of culturally relevant fungi in Mexico has been recorded in temperate forests. Likewise, it has been proven that people who live in such environments possess greater ethnomycological knowledge, compared with people in the lowlands. In this study, we investigated whether the uses and perceptions of fungi were different between people living near forests in highlands and those living in lowlands near grasslands and subtropical scrublands. We selected seven communities from the municipality of Tlaltenango, Zacatecas, Mexico, where we conducted guided tours, 35 semistructured interviews, and free lists to 420 people. Since highlands surrounded by temperate forests give rise to high species diversity, we expected that the inhabitants, in comparison with those of the lowlands where there is less diversity, would recognize a larger number of fungi. To test this hypothesis, we employed discriminant function analysis, principal coordinate analysis, Mann-Whitney U test, and linear regression. The cultural importance of each species was calculated based on the frequency with which it was mentioned and the first principal coordinate. Although the highlanders have a deeper knowledge of mushrooms, lowlanders and highlanders had similar fungal backgrounds and preferred the same species, regardless of the fungi surrounding their territory. The lack of differences among communities was due in part to the fact that the most culturally important species were those that grew in grasslands and subtropical scrub areas. Agaricus campestris was the most culturally prevalent and only commercialized species both in the highlands and lowlands, followed by Pleurotus djamor and Volvariella bombycina. None of the mushrooms growing in the pine-oak forest had a high cultural importance value, even for the communities living in the vicinity of this forest, forcing them to travel long distances to collect Agaricus campestris. Further investigations are needed in order to assess the relevance of sociocultural factors, and their potential influence in the preference for particular mushrooms in this region.