Termite Mushrooms (Termitomyces), a Potential Source of Nutrients and Bioactive Compounds Exhibiting Human Health Benefits: A Review.

Termite mushrooms have been classified to the genus Termitomyces, family Lyophyllaceae, order Agaricales. These mushrooms form a mutualistic association with termites in the subfamily Macrotermitinae. In fact, all Termitomyces species are edible and have unique food value attributed to their texture, flavour, nutrient content, and beneficial mediational properties. Additionally, Termitomyces have been recognized for their ethno-medicinal importance in various indigenous communities throughout Asia and Africa. Recent studies on Termitomyces have indicated that their bioactive compounds have the potential to fight against certain human diseases such as cancer, hyperlipidaemia, gastroduodenal diseases, and Alzheimer's. Furthermore, they possess various beneficial antioxidant and antimicrobial properties. Moreover, different enzymes produced from Termitomyces have the potential to be used in a range of industrial applications. Herein, we present a brief review of the current findings through an overview of recently published literature involving taxonomic updates, diversity, distribution, ethno-medicinal uses, nutritional value, medicinal importance, and industrial implementations of Termitomyces, as well as its socioeconomic importance.

Ganoderma (Ganodermataceae, Basidiomycota) Species from the Greater Mekong Subregion.

The cosmopolitan fungal genus Ganoderma is an important pathogen on arboreal plant hosts, particularly in tropical and temperate regions. It has long been used as a traditional medicine because of its medicinal properties and chemical constituents. In this study, Ganoderma collections were made in the Greater Mekong Subregion (GMS), encompassing tropical parts of Laos, Myanmar, Thailand, Vietnam, and temperate areas in Yunnan Province, China. The specimens used in this study are described based on micro-macro-characteristics and phylogenetic analysis of combined ITS, LSU, TEF1α, and RPB2 sequence data. In this comprehensive study, we report 22 Ganoderma species from the GMS, namely, G. adspersum, G. applanatum, G. australe, G. calidophilum, G. ellipsoideum, G. flexipes, G. gibbosum, G. heohnelianum, G. hochiminhense, G. leucocontextum, G. lucidum, G. multiplicatum, G. multipileum, G. myanmarense, G. orbiforme, G. philippii, G. resinaceum, G. sichuanense, G. sinense, G. subresinosum, G. williamsianum, and G. tsugae. Some of these species were reported in more than one country within the GMS. Of these 22 species, 12 were collected from Yunnan Province, China; three were collected from Laos; three species, two new records, and one new species were collected from Myanmar; 15 species and four new records were collected from Thailand, and one new species was collected from Vietnam. Comprehensive descriptions, color photographs of macro- and micro-characteristics, the distribution of Ganoderma within the GMS, as well as a phylogenetic tree showing the placement of all reported Ganoderma from the GMS are provided.

Using High-Throughput Amplicon Sequencing to Evaluate Intragenomic Variation and Accuracy in Species Identification of Cordyceps Species.

While recent sequencing technologies (third generation sequencing) can successfully sequence all copies of nuclear ribosomal DNA (rDNA) markers present within a genome and offer insights into the intragenomic variation of these markers, high intragenomic variation can be a source of confusion for high-throughput species identification using such technologies. High-throughput (HT) amplicon sequencing via PacBio SEQUEL I was used to evaluate the intragenomic variation of the ITS region and D1-D2 LSU domains in nine Cordyceps species, and the accuracy of such technology to identify these species based on molecular phylogenies was also assessed. PacBio sequences within strains showed variable level of intragenomic variation among the studied Cordyceps species with C. blackwelliae showing greater variation than the others. Some variants from a mix of species clustered together outside their respective species of origin, indicative of intragenomic variation that escaped concerted evolution shared between species. Proper selection of consensus sequences from HT amplicon sequencing is a challenge for interpretation of correct species identification. PacBio consensus sequences with the highest number of reads represent the major variants within a genome and gave the best results in terms of species identification.

An untold story of a novel mushroom from tribal cuisine: an ethno-medicinal, taxonomic and pharmacological approach.

India showcases an array of fascinating and rare mushrooms that grow exclusively in the wilderness of West Bengal. Thus, the state has always been our prime choice to document myco-diversity and associated indigenous knowledge. Fortuitously, a recent expedition gifted us a violet-coloured Russuloid macrofungus, called "Jam Patra", that plays an integral part in the food security of local ethnic groups. However, the species has not received the much-needed attention among city dwellers and remains abandoned, motivating us to carry a thorough investigation. To our surprise, extensive analyses on morphological features and nrITS based phylogenetic estimation pointed the novelty of the taxon, as justified herein. Extending this research, a water-soluble polysaccharide-rich fraction was isolated to determine therapeutic prospects. Chemical characterization revealed that the backbone of the polymers, organized in triple-helical form, predominantly consisted of ß-glucan accompanied by a lower extent of galactose, mannose and xylose. Subsequently, the effective antioxidant activity was noted in terms of radical scavenging, reducing power and chelating ability with EC50 of 305-2726 µg ml-1. Further, the macromolecules triggered murine macrophages to proliferate, phagocytose, release NO, produce intracellular ROS and change morphodynamics. A significant alleviation in the expression of TLR-2, TLR-4, NF-κB, COX-2, TNF-α, Iκ-Bα, IFN-γ, IL-10 and iNOS was also observed explaining the definite immune-stimulatory activity and supporting traditional consumption of "Jam Patra" as a health-promoting food. Altogether, the study introduces a species in the world's myco-diversity and tribal food list opening doors of various opportunities in functional food and nature-based drug development arenas, which are currently in trend.

Selective in vitro inhibition of Leishmania donovani by a semi-purified fraction of wild mushroom Grifola frondosa.

The current study was designed to assess the anti-leishmanial effect of a semi-purified fraction of wild mushroom Grifola frondosa against Leishmania donovani, in vitro. A total of five extracts from three wild mushrooms [Grifola frondosa (family, Meripilaceae) Laetiporus sulphurous (family, Polyporaceae) and Meripilus giganteus (family, Meripilaceae) were explored for novel anti-leishmanial leads against promastigotes. The ethanol extract of G. frondosa was selected as the most efficient against L. donovani promastigotes (IC50: 93.9 µg/mL). A semi-purified fraction was obtained from an active ethanol extract of G. frondosa and found to inhibit the survival of promastigotes of L. donovani (MHOM/IN/83/AG83) significantly (IC50: 20.37 µg/mL) and it also had some effect against L. major LV39 (MRHO/Sv/59/P strain) and L. tropica WR683 (MHOM/SU/58/OD) strains at higher concentrations (IC50: 46.08 µg/mL and 53.79 µg/mL respectively). The semi-purified fraction also interfered in lipid biosynthesis, altered parasite morphology and induced apoptosis in L. donovani promastigotes. The semi-purified fraction was also effective against intracellular amastigotes in infected macrophages and enhanced the release of nitric oxide and pro-inflammatory cytokines, in vitro. Interestingly, the 50% inhibitory concentration of the semi-purified fraction against the intracellular amastigotes (IC50: 2.48 µg/mL) was much lower in comparison to promastigotes (IC50: 20.37 µg/mL). The semi-purified fraction was found to inhibit the intracellular amastigotes slightly more efficiently in comparison to conventional anti-leishmanial drugs; sodium antimony gluconate, amphotericin B, miltefosine and paromomycin and noticeably non-toxic towards host splenocytes. The findings of the present study established that G. frondosa might be a natural resource for development of a new anti-leishmanial lead.

Introducing a novel mushroom from mycophagy community with emphasis on biomedical potency.

Mushrooms have been prized by humankind as medicine and culinary wonder since antiquity. Though several species are ethnically valued; many prospective species are still being discovered. One such wild macrofungus has recently been discovered during subsequent field surveys in West Bengal, India which in turn exposed as a traditionally consumed popular myco-food. The collected taxon was found to be unique with regard to its morphological as well as genetical features. After detailed characterizations, the fungus was identified as a novel taxon belonging to the genus Russula (Russulaceae, Basidiomycota). Besides, the investigation was further extended in search of new functional ingredients and in this context, a water soluble crude polysaccharide rich extract (Rusalan) was isolated from dried basidiocarps. Accumulating evidences from GC-MS, HPTLC, FT-IR along with several spectrophotometric methods postulated that the fraction consisted mainly of carbohydrate in triple helical conformation, where glucose was the major monosaccharide mostly with ß-type glycosidic linkage. Conversely, Rusalan showed pronounced antioxidant activity in six in vitro assay systems with EC50 value ranging from 190-1328 µg/ml concentration. The crude polysaccharide was also evaluated against six bacterial strains using microdilution method and the growth of Staphylococcus aureus and Bacillus subtilis were found to be inhibited effectively. In addition, immune-stimulatory assays demonstrated that Rusalan could evidently promote proliferation, induce phagocytosis, release NO, produce intracellular ROS and upregulate mRNA expression of iNOS, TNF-α, COX-2, as well as IL-6 genes in in mouse macrophage cells. Therefore, aim of the present study was not only to describe a new taxon to the world mycoflora but also to introduce a potent therapeutic agent that could be explored for food and pharmaceutical purposes. However, isolation of active component and in vivo studies need to be designed further.

Structural characterization and antioxidant activity of a glucan from Meripilus giganteus.

A new water soluble glucan (MGPS), with a molecular weight ∼1.48×105Da, was isolated from the fruit bodies of Meripilus giganteus by hot water extraction followed by purification through dialysis tubing cellulose membrane and sepharose 6B column chromatography. Its structural characteristics were investigated by acid hydrolysis, methylation analysis, Smith degradation and 1D/2D NMR experiments. The monosaccharide composition analysis showed that MGPS contain only glucose. The backbone of MGPS was composed of two (1→3)-ß-d-glucopyranosyl, two (1→6)-ß-d-glucopyranosyl, two (1→6)-α-d-glucopyranosyl, and one (1→4)-α-d-glucopyranosyl residues, out of which one (1→3)-ß-d-glucopyranosyl residue was branched at O-6 position with terminal α-d-glucopyranosyl residue and one (1→4)-α-d-glucopyranosyl residue branched at O-6 position with terminal ß-d-glucopyranosyl residue. In vitro antioxidant studies showed that the MGPS exhibited hydroxide radical scavenging activity (IC50=390µg/mL), superoxide radical scavenging activity (IC50=70µg/mL), and ferrous ion chelating activity (IC50=290µg/mL).

Structural, immunological, and antioxidant studies of ß-glucan from edible mushroom Entoloma lividoalbum.

A water soluble ß-glucan having molecular weight ∼2×10(5)Da was isolated from hot water extract of the fruit bodies of an edible mushroom Entoloma lividoalbum (Kühner & Romagn) Kubicka. This polysaccharide (ELPS) contains (1→3,6)-ß-D-Glcp, (1→3)-ß-D-Glcp, (1→6)-ß-D-Glcp, and terminal ß-D-Glcp moieties in a molar ratio of nearly 1:1:3:1. Chemical and spectroscopic analysis showed that the backbone of glucan consists of three (1→6)-ß-D-glucopyranosyl and two (1→3)-ß-D-glucopyranosyl residues, out of which one (1→3)-ß-D-glucopyranosyl moiety was branched at O-6 with a terminal ß-D-glucopyranosyl residue. This ß-glucan exhibited macrophage, splenocyte, and thymocyte stimulations. It possesses promising antioxidant activities as evidenced from its hydroxyl and superoxide radical scavenging activities and reducing properties.

Selective inhibition of Leishmania donovani by active extracts of wild mushrooms used by the tribal population of India: An in vitro exploration for new leads against parasitic protozoans.

The study was intended at evaluating the anti-proliferating effect of mushrooms used in traditional folklore of Santal tribal population in India against Leishmania donovani (MHOM/IN/83/AG83). A total of eighteen extracts, three estracts from each mushroom [(80% ethanol extracted; Fa), (water-soluble polysaccharide fraction; Fb), (polyphenolic fraction; Fc)], from six wild mushrooms were obtained. These extracts were tested against the promastigotes and amastigotes for their antileishmanial capacity. Fa fractions (250 µg/mL) of Astraeus hygrometricus and Tricholoma giganteum significantly inhibited the growth of L. donovani promastigotes and interfered in lipid biosynthesis. Moreover, both fractions induced apoptosis in promastigotes. Water soluble Fb fractions of A. hygrometricus, Russula laurocerasi, Russula albonigra, Termitomyces eurhizus, Russula delica and polyphenolic Fc fraction of R. laurocerasi were found to inhibit the replication of intracellular amastigotes in macrophages dose dependently. Significantly, 50% inhibitory concentration of the active extracts against intracellular amastigotes induced release of nitric oxide and IL-12 in murine macrophages and dendritic cells assay and also found considerably non-toxic on murine splenocytes. Results of this study can be used as a basis for further phytochemical and pharmacological investigations in the effort for search of novel anti-leishmanial leads.

Structure elucidation and antioxidant properties of a soluble ß-D-glucan from mushroom Entoloma lividoalbum.

A water soluble branched ß-D-glucan (PS-I) with an average molecular weight ~2.1×10(5) Da was isolated from alkaline extract of the fruit bodies of the edible mushroom Entoloma lividoalbum (Kühner & Romagn) Kubicka which consists of terminal ß-D-glucopyranosyl, (1→3)-ß-D-glucopyranosyl, (1→6)-ß-D-glucopyranosyl, and (1→3,6)-ß-D-glucopyranosyl moieties in a molar ratio of nearly 1:3:2:1. The structure of PS-I was elucidated using acid hydrolysis, methylation analysis, periodate oxidation study, partial hydrolysis, and 1D/2D NMR experiments. The repeating unit of the polysaccharide (PS-I) contains a backbone chain of three (1→6)-ß-D-glucopyranosyl residues, one of which was branched at O-3 position with the side chain consisting of three (1→3)-ß-D-glucopyranosyl and a terminal ß-D-glucopyranosyl residues. Total antioxidant capacity of 1mg PS-I was measured and found equivalent to 70±15 µg of ascorbic acid. The PS-I was found to possess hydroxyl and superoxide radical-scavenging activities with EC50 values of 480 and 150 µg/mL, respectively. The reducing power of PS-I was determined 0.5 at 480 µg/mL.

Polysaccharide-rich fraction of Termitomyces eurhizus accelerate healing of indomethacin induced gastric ulcer in mice.

The current study aims to determine the healing activity of water soluble polysaccharide-rich fraction of a wild mushroom, Termitomyces eurhizus (TEps) against the indomethacin induced gastric ulceration in mice model. Gastric tissue histology, myeloperoxidase (MPO) activity, cyclooxygenases (COX) 1 and 2 expression, prostaglandin E2 (PGE2) synthesis, and modulation of pro/anti inflammatory cytokines expression were studied for this purpose. Histological study shows that TEps (20 mg/kg) effectively healed the gastric ulceration. Based on biochemical results, the healing capacities of TEps could be attributed to reduction of MPO activity and protection of mucosal mucin content. Enhanced synthesis of PGE2 by modulation of COX-1 and COX-2 expression and a prominent shift of cytokines expression from pro (TNF-α, IL-1ß) to anti inflammatory (IL-10) side are also held responsible for ulcer healing. The preliminary study highlights the anti-ulcerogenic property of polysaccharide-rich fraction of Termitomyces eurhizus and opens an alternative cure for NSAID induced gastroduodenal diseases.