Array of Metabolites in Italian Hericium erinaceus Mycelium, Primordium, and Sporophore.

Hericium erinaceus is a medicinal mushroom that contains many molecules promising a plethora of therapeutic properties. In this study, the strain H.e.2 (MicUNIPV, University of Pavia, Italy) was isolated from a sporophore collected in Tuscany (Italy). Mycelium, primordium, and wild type and cultivated sporophores were analyzed by HPLC-UV-ESI/MS. Erinacine A in the mycelium and hericenones C and D in the sporophores were quantified by comparison with their standard molecules. For the first time, H. erinaceus primordium was also investigated for the presence of these molecules. Comparing with the literature data, hericenes, molecules structurally similar to hericenones, were present in all our samples. The highest contents of hericenones C and D were detected in cultivated sporophores, compared to the wild type. The comparison of these data with those of another Italian H. erinaceus strain (H.e.1 MicUNIPV) was discussed. The results led us to select H. erinaceus strains more suitable for mycelium production or sporophore cultivation to obtain extracts with a higher content of bioactive compounds. This work provides a further step towards standardizing the procedures in the development of dietary supplements made from mushrooms.

Ganoderma resinaceum and Perenniporia fraxinea: Two Promising Wood Decay Fungi for Pharmaceutical Degradation.

Wood decay fungi (WDF) are a well-known source of enzymes and metabolites which have applications in numerous fields, including myco-remediation. Pharmaceuticals are becoming more problematic as environmental water pollutants due to their widespread use. In this study, Bjerkandera adusta, Ganoderma resinaceum, Perenniporia fraxinea, Perenniporia meridionalis and Trametes gibbosa were chosen from WDF strains maintained in MicUNIPV (the fungal research collection of the University of Pavia) to test their potential to degrade pharmaceuticals. The degradation potential was tested in spiked culture medium on diclofenac, paracetamol and ketoprofen, three of the most common pharmaceuticals, and irbesartan, a particularly difficult molecule to degrade. G. resinaceum and P. fraxinea were found to be the most effective at degradation, achieving 38% and 52% (24 h) and 72% and 49% (7 d) degradations of diclofenac, 25% and 73% (24 h) and 100% (7 d) degradations of paracetamol and 19% and 31% (24 h) and 64% and 67% (7 d) degradations of ketoprofen, respectively. Irbesartan was not affected by fungal activity. The two most active fungi, G. resinaceum and P. fraxinea, were tested in a second experiment in discharge wastewater collected from two different wastewater treatment plants in northern Italy. A high degradation was found in azithromycin, clarithromycin and sulfametoxazole (from 70% up to 100% in 7 days).

Diversity of Pleurotus spp. (Agaricomycetes) and Their Metabolites of Nutraceutical and Therapeutic Importance.

Pleurotus spp. have been gaining popularity as a source for the creation of functional foods, nutraceuticals and novel pharmaceuticals. Despite Pleurotus is a specious genus including 208 legitimate species, only a few of them such as P. ostreatus are commercially accessible. The genetic and metabolic diversity of Pleurotus both at specific and subspecific level is therefore of main concern for many researchers. In addition to the conventional morphological approach, molecular and biochemical markers have been greatly contributing to investigate these issues. In this study, samples from six Pleurotus species (P. eryngii is represented by three varieties) were molecularly identified and the phylogeny was inferred to assess the relationships between the various taxa. Strains in pure culture obtained from 6 out of 7 species were cultivated as mycelium in vitro to investigate the metabolites by untargeted LC-MS/MS-based metabolomics. The results pointed out species-specific metabolite patterns and highlighted a clear difference between the P. eryngii group and P. ostreatus, although the latter appears more versatile depending on the strain. This is the first study pointing out and comparing different metabolite patterns in Italian samples of Pleurotus species, including P. eryngii varieties.

The Analysis of the Mycobiota in Plastic Polluted Soil Reveals a Reduction in Metabolic Ability.

Plastic pollution is a growing environmental issue that results in its accumulation and persistence in soil for many decades, with possible effects on soil quality and ecosystem services. Microorganisms, and especially fungi, are a keystone of soil biodiversity and soil metabolic capacity. The aim of this research was to study soil fungal biodiversity and soil microbial metabolic profiles in three different sites in northern Italy, where macro- and microplastic concentration in soil was measured. The metabolic analyses of soil microorganisms were performed by Biolog EcoPlates, while the ITS1 fragment of the 18S ribosomal cDNA was used as a target for the metabarcoding of fungal communities. The results showed an intense and significant decrease in soil microbial metabolic ability in the site with the highest concentration of microplastics. Moreover, the soil fungal community composition was significantly different in the most pristine site when compared with the other two sites. The metabarcoding of soil samples revealed a general dominance of Mortierellomycota followed by Ascomycota in all sampled soils. Moreover, a dominance of fungi involved in the degradation of plant residues was observed in all three sites. In conclusion, this study lays the foundation for further research into the effect of plastics on soil microbial communities and their activities.

Fungal Diversity in Two Wastewater Treatment Plants in North Italy.

In urban wastewater treatment plants, bacteria lead the biological component of the depuration process, but the microbial community is also rich in fungi (mainly molds, yeasts and pseudo-yeasts), whose taxonomical diversity and relative frequency depend on several factors, e.g., quality of wastewater input, climate, seasonality, and depuration stage. By joining morphological and molecular identification, we investigated the fungal diversity in two different plants for the urban wastewater treatment in the suburbs of the two major cities in Lombardia, the core of industrial and commercial activities in Italy. This study presents a comparison of the fungal diversity across the depuration stages by applying the concepts of α-, ß- and ζ-diversity. Eurotiales (mainly with Aspergillus and Penicillium), Trichosporonales (Trichosporon sensu lato), Saccharomycetales (mainly with Geotrichum) and Hypocreales (mainly with Fusarium and Trichoderma) are the most represented fungal orders and genera in all the stages and both the plants. The two plants show different trends in α-, ß- and ζ-diversity, despite the fact that they all share a crash during the secondary sedimentation and turnover across the depuration stages. This study provides an insight on which taxa potentially contribute to each depuration stage and/or keep viable propagules in sludges after the collection from the external environment.

Untargeted Metabolomics Used to Describe the Chemical Composition, Antioxidant and Antimicrobial Effects of Extracts from Pleurotus spp. Mycelium Grown in Different Culture Media.

Pleurotus species isolated in vitro were studied to determine the effect of different media on their production of secondary metabolites, antimicrobial, and antioxidant activity. The different metabolites among Pleurotus samples covered a total of 58 pathways. Comparisons were made between the metabolic profiles of Pleurotus spp. mycelia grown in two substrates: Potato-dextrose-agar-PDA, used as control (S1), and PDA enriched with 0.5 % of wheat straw (S2). The main finding was that the metabolic pathways are strongly influenced by the chemical composition of the growth substrate. The antibacterial effects were particularly evident against Escherichia coli, whereas Arthroderma curreyi (CCF 5207) and Trichophyton rubrum (CCF 4933) were the dermatophytes more sensitive to the mushroom extracts. The present study supports more in-depth investigations, aimed at evaluating the influence of growth substrate on Pleurotus spp. antimicrobial and antioxidant properties.

White-Nose Syndrome Confirmed in Italy: A Preliminary Assessment of Its Occurrence in Bat Species.

Although no mass mortality has been recorded so far, the precise demographic effect of white-nose syndrome (WNS) on European bats still remains to be ascertained. Following the first isolation of P. destructans in Italy, further surveys were performed to assess the distribution of the fungus in NW Italy and its effects on bats. Data were collected from March 2019 to April 2020 at sites used for hibernation (six sites) and/or for reproduction (four sites) in Piedmont and Aosta Valley. A total of 138 bats, belonging to 10 species, were examined to identify clinical features possibly related to the fungal presence. Culture from swabs and the molecular identification of isolates confirmed the presence of P. destructans in bats from five sites, including two maternal roosts. Dermal fungal infiltration, the criterion to assess the presence of WNS, was observed in biopsies of bats belonging to Myotis blythii, M. daubentonii, M. emarginatus and M. myotis. This is the first report of the disease in Italy. The results suggest a greater susceptibility to the infection of the genus Myotis and particularly of M. emarginatus, possibly due to the long length of its hibernation period. Other fungal dermatophytes were also observed.

Collection and Characterization of Wood Decay Fungal Strains for Developing Pure Mycelium Mats.

Wood decay fungi (WDF) seem to be particularly suitable for developing myco-materials due to their mycelial texture, ease of cultivation, and lack of sporification. This study focused on a collection of WDF strains that were later used to develop mycelium mats of leather-like materials. Twenty-one WDF strains were chosen based on the color, homogeneity, and consistency of the mycelia. The growth rate of each strain was measured. To improve the consistency and thickness of the mats, an exclusive method (newly patented) was developed. The obtained materials and the corresponding pure mycelia grown in liquid culture were analyzed by both thermogravimetric analysis (TGA) and scanning electron microscopy (SEM) to evaluate the principal components and texture. TGA provided a semi-quantitative indication on the mycelia and mat composition, but it was hardly able to discriminate differences in the production process (liquid culture versus patented method). SEM provided keen insight on the mycelial microstructure as well as that of the mat without considering the composition; however, it was able to determine the hyphae and porosity dimensions. Although not exhaustive, TGA and SEM are complementary methods that can be used to characterize fungal strains based on their desirable features for various applications in bio-based materials. Taking all of the results into account, the Fomitopsis iberica strain seems to be the most suitable for the development of leather-like materials.

Antimicrobial properties of Fomitopsis officinalis in the light of its bioactive metabolites: a review.

Fomitopsis officinalis, also known as Laricifomes officinalis, is a medicinal polypore used for millennia (Agarikon) to contrast several diseases, particularly the pulmonary ones. A rich literature has dealt with its ethno-mycological aspects, but isolation and chemical characterisation of single compounds has only recently significantly developed, as well as in vitro tests for bioactivity. According to several reports there is evidence of a broad-spectrum antibacterial and antiviral activity by F. officinalis, including pathogens like Mycobacterium tuberculosis, Yersinia pseudotuberculosis and Staphylococcus aureus, as well as Ortopox virus. Chlorinated coumarins from mycelia and lanostane triterpenoids from basidiomes have been demonstrated to be directly responsible for antiviral-antibacterial and trypanocidal activity, respectively. A wider literature deals instead with crude extracts including an undetermined mixture of metabolites, whose efficacy in vitro is yet far from being standardised as extraction and treatment methodology are highly variable. Nevertheless, in vivo tests on bees provided promising results in order to develop sustainable solutions against the pathogens responsible for colony collapse disorders. Despite increasing attention has been paid to other medicinal aspects of this polypore, such as immune-tropic or antitumor, this review rationally reports and critically analyses the available knowledge by focusing on aspects of antimicrobial properties.

Efficacy of bergamot: From anti-inflammatory and anti-oxidative mechanisms to clinical applications as preventive agent for cardiovascular morbidity, skin diseases, and mood alterations.

We summarize the effects of bergamot (extract, juice, essential oil, and polyphenolic fraction) on cardiovascular, bone, inflammatory, skin diseases, mood alteration, anxiety, pain, and stress. This review included a total of 31 studies (20 studies on humans with 1709 subjects and 11 in animals (rats and mice)). In humans, bergamot-derived extract (BE) exerts positive effects on hyperlipidemia with an oral dose from 150 mg to 1000 mg/day of flavonoids administered from 30 to 180 days, demonstrating an effect on body weight and in modulating total cholesterol, triglycerides, LDL, and HDL. Studies in animals confirm promising data on glucose control (500/1000 mg/day of BE with a treatment lasting 30 days) are available in rats. In animals models, bergamot essential oil (BEO, 10 mg/kg or 20 mg/kg daily for 20 weeks) increases bone volume, decreases psoriatic plaques, increases skin collagen content, and promotes hair growth. Bergamot juice (20 mg/kg) is promising in terms of pro-inflammatory cytokine reduction. In humans, aromatherapy (from 15 to 30 min) does not appear to be useful in order to reduce stress, anxiety, and nausea, compared to placebo. Compared to baseline, BE topical application and BEO aromatherapy reduce blood diastolic and systolic pressure and could have a significant effect on improving mental conditions.

Phylogenetic Comparison between Italian and Worldwide Hericium Species (Agaricomycetes).

A broad literature concerns the genus Hericium, mainly regarding the medicinal properties of H. erinaceus. Congeneric species of H. erinaceus have been poorly investigated. We collected basidiomata of H. alpestre, H. coralloides and H. erinaceus in Italy and isolated the corresponding mycelia in pure culture. Analysis of the respective internal transcribed spacer regions confirmed the morphological identification of the strains. Internal transcribed spacer sequences from the Italian strains were phylogenetically compared along with 64 other sequences available from Gen-Bank, the CBS Strain Database, and the European Nucleotide Archive (ENA) for the same Hericium . Geographic origin and host plant species were cross-checked using the above data banks. Bayesian phylogenetic analysis produced a phylogram that permitted good discrimination among Hericium species. It provides an updated phylogeny within the genus Hericium and a better understanding of affinity among the species analyzed. The main Hericium clade includes the following: the H. erinaceus group and the H. alpestre/H. coralloides group, where the two species cluster separately. This study also allowed us to differentiate the H. erinaceus group on a biogeographical basis. The phylogenetic comparison further confirms the importance of a joint morphological-molecular approach to avoid misidentification and to guarantee the quality of strains for further chemical and medicinal characterization.

Hericium erinaceus Improves Mood and Sleep Disorders in Patients Affected by Overweight or Obesity: Could Circulating Pro-BDNF and BDNF Be Potential Biomarkers?

Epidemiological data indicate that subjects affected by obesity have an increased risk of developing mood disorders. The relationship between obesity and mood disorders is bidirectional. We assessed whether a Hericium erinaceus treatment improved depression, anxiety, sleep, and binge eating disorders after 8 weeks of supplementation in subjects affected by overweight or obesity under a low calorie diet regimen. Looking for a possible clinical biomarker, we assessed the serum balance between brain-derived neurotrophic factor (BDNF) and its precursor pro-BDNF before and after H. erinaceus supplementation. Seventy-seven volunteers affected by overweight or obesity were recruited at the offices of the Department of Preventive Medicine, Luigi Devoto Clinic of Work, Obesity Centre, at the IRCCS Foundation Policlinico Hospital of Milan (Italy). Patients were recruited only if they had a mood and/or sleep disorder and/or were binge eating as evaluated through self-assessment questionnaires. We used two different enzyme-linked immunosorbent assays kits to discriminate circulating levels of pro-BDNF and BDNF. Eight weeks of oral H. erinaceus supplementation decreased depression, anxiety, and sleep disorders. H. erinaceus supplementation improved mood disorders of a depressive-anxious nature and the quality of the nocturnal rest. H. erinaceus increased circulating pro-BDNF levels without any significant change in BDNF circulating levels.

Hericium erinaceus Improves Recognition Memory and Induces Hippocampal and Cerebellar Neurogenesis in Frail Mice during Aging.

Frailty is a geriatric syndrome associated with both locomotor and cognitive decline, implicated in both poor quality of life and negative health outcomes. One central question surrounding frailty is whether phenotypic frailty is associated with the cognitive impairment during aging. Using spontaneous behavioral tests and by studying the dynamic change during aging, we demonstrated that the two form of vulnerability, locomotor and recognition memory decline, develop in parallel and therefore, integration of the motoric and cognitive evaluations are imperative. We developed an integrated frailty index based on both phenotypic and recognition memory performances. Hericium erinaceus (H. erinaceus) is a medicinal mushroom that improves recognition memory in mice. By using HPLC-UV-ESI/MS analyses we obtained standardized amounts of erinacine A and hericenones C and D in H. erinaceus extracts, that were tested in our animal model of physiological aging. Two-month oral supplementation with H. erinaceus reversed the age-decline of recognition memory. Proliferating cell nuclear antigen (PCNA) and doublecortin (DCX) immunohistochemistry in the hippocampus and cerebellum in treated mice supported a positive effect of an H. erinaceus on neurogenesis in frail mice.

Dietary Supplementation of Lion's Mane Medicinal Mushroom, Hericium erinaceus (Agaricomycetes), and Spatial Memory in Wild-Type Mice.

Hericium erinaceus is an edible and medicinal mushroom with potential neuroprotective effects. The study of H. erinaceus has attracted considerable attention during the past 10 years, particularly with regard to its potential utility in the treatment of motor dysfunction, Alzheimer disease, and other forms of dementia. We previously determined that oral supplementation with H. erinaceus results in significant improvements in novelty-seeking behavior and novel object recognition in mice. In this study, H. erinaceus was added to the diets of wild-type mice for 2 months, and effects on spatial memory were evaluated by means of a Y maze and an object location task. We found that H. erinaceus increased general locomotor activity but had no effect on spatial memory. Thus, oral supplementation with H. erinaceus yields specific and selective improvements in recognition memory without altering spatial working memory, which supports the hypothesis that recognition memory can be modeled as a dual process. In this model, the perirhinal cortex supports the recognition of individual items as part of a circuit involved in familiarity with an encountered stimulus, whereas the hippocampus supports recollected associations and relationships between stimuli.

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

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