Toxicity, mutagenicity and trace metal constituent of Termitomyces schimperi (Pat.) R. Heim (Lyophyllaceae) and kaolin, a recipe used traditionally in cancer management in Cote d'Ivoire.

ETHNOPHARMACOLOGICAL RELEVANCE: Some local communities in Cote d'Ivoire use the mushroom Termitomyces schimperi combined with kaolin (TSK) to manage various cancers in patients. However, there is a paucity of data on toxicity, mutagenicity and trace metal constituent of TSK. AIM OF THE STUDY: We sought to investigate the acute and sub-chronic toxicities, mutagenic potential, and trace metal constituents of TSK. MATERIALS AND METHODS: To assess acute toxicity, single doses (1000, 3000 and 5000 mg/kg) of aqueous extract of TSK were administrated per os to Sprague Dawley (SD) rats on Day 1. The rats were then monitored for 13 consecutive days. Sub-chronic toxicity was evaluated by daily administration of 200 and 500 mg/kg of the extract per os for 90 consecutive days. SD rats used as control received distilled water. Signs of toxicity, changes in body weight and mortality were monitored. After the aforementioned monitoring processes, rats were sacrificed and blood collected for full blood count and biochemistry analysis. Animal organs were also collected for histopathological examination. The mutagenic potential of the aqueous extract of TSK (10000 µg/mL) on TA98 Salmonella typhimurium was estimated. Additionally, energy-dispersive X-ray fluorescence (ED-XRF) method was employed to determine trace metal constituents of TSK. RESULTS: Single-dose administration of 5000 mg/kg of TSK did not cause any death in the SD rats; thus, LD50 was above 5000 mg/kg. Administration of 1000 and 3000 mg/kg of the aqueous extract of TSK did not cause any significant change in behaviour and body weight of SD rats during the 14-day monitoring period. However, the mean corpuscular volume and the mean corpuscular haemoglobin concentration increased significantly (p < 0.01) among rats administered 1000 and 3000 mg/kg of TSK. There was a significant increase (p < 0.0001) in alanine transaminase levels in rats administered 1000 and 3000 mg/kg of TSK extract compared with control. Conversely, there was a significant decrease (p=0.0122) in serum creatine level among rats administered 1000 and 3000 mg/kg of TSK extract compared with control. After 14 days, there were minimal changes with isolated organs of TSK-treated and control rats. Furthermore, 90-day treatment with extract of TSK caused no significant change in parameters assessed. TSK induced frameshift gene mutation in S. typhimurium before (p < 0.05) and after metabolic activation (p < 0.001). Elemental analysis of TSK revealed the presence of toxic (aluminium) or potentially toxic (silver, rabidium, titanium and zirconium) elements. CONCLUSIONS: The aqueous extract of TSK showed no toxicity (acute and sub-chronic) at doses tested. These findings are consistent with the absence of heavy metals (i.e., cadmium) and potentially toxic elements (i.e., uranium) in TSK samples analysed. TSK showed some level of mutagenic potential. Further mutagenic and chronic toxicity studies on TSK are required.

Investigation of Antioxidant and Antimicrobial Activities of Different Extracts of Auricularia and Termitomyces Species of Mushrooms.

Mushrooms produce a variety of bioactive compounds that are known to have a potential source of antioxidant and antimicrobial properties. Natural antioxidants can protect against free radicals without any side effects. The purpose of this study was to evaluate the antioxidant and antimicrobial activities of Auricularia and Termitomyces extracts. Specimens of Auricularia and Termitomyces spp. were collected from Kakamega National Reserve Forest in Kenya. Specimens were identified, extracted, and screened for their antioxidant and antimicrobial activities using stable free radical DPPH and colorimetric bioassay methods, respectively. The antimicrobial activity of the extracts was tested against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, MRSA, Candida albicans, and Candida parapsilosis. The maximum scavenging activity of hot water extract of Auricularia spp. was observed at 70.4% with the IC50 value of 40 µg/mL. Of the three extracts of Termitomyces spp., 70% ethanol extract has shown the highest scavenging activity (63%) with the IC50 value of 50 µg/mL. Chloroform and hot water extracts of Auricularia have shown statistically significantly different antifungal activities against C. parapsilosis (df = 2, F = 22.49, p ≤ 0.05). Of all the organisms, S. aureus was highly susceptible to 70% ethanol and hot water extracts of Termitomyces spp. with minimum inhibitory concentration values of 0.67±0.29 mg/mL. S. aureus and E. coli were the most susceptible and resistant bacteria to the hot water extract, respectively. In conclusion, the extracts of Auricularia spp. and Termitomyces spp. have shown promising antimicrobial and antioxidant activities.

Safety Evaluation of an Aqueous Extract of Termitomyces robustus (Agaricomycetes) in Wistar Rats.

Termitomyces robustus is an edible and highly nutritious wild Basidiomycetes mushroom. It is used in ethnomedicine for treating malnutrition-related diseases, rheumatism, diarrhea, gonorrhea, anemia, and hypertension. Despite the tremendous use of this delicious edible mushroom as a source of nutrients, no comprehensive literature describes its safety and toxicity profiles. Therefore, this study evaluated the toxicity profile of an aqueous T. robustus extract in rats. In the acute toxicity test, male and female rats were orally administered daily a single dose of up to 10 g/kg extract. In the subacute toxicity test, male rats were orally administered the T. robustus extract at graded doses of 500, 1000, and 1500 mg/kg for 14 days. No mortality or any signs of toxicity were observed in the acute toxicity study, indicating that the median lethal dose (LD50) of T. robustus is greater than 10 g/kg. In the subacute toxicity study, T. robustus had no effect (P > 0.05) on hemoglobin, packed cell volume, red blood cell, white blood cell, alanine aminotransferase, alkaline phosphatase, neutrophil, lymphocyte, or lipid profile parameters in any of the rats. However, significant differences (P < 0.05) were noted in alanine aminotransferase, eosinophils, basophils, monocytes, platelets, urea, creatinine, and electrolytes in the tested groups when compared to values from the control group. No histopathological alterations or changes were observed in the liver or kidneys of the rats. This study established that an aqueous extract of T. robustus is nontoxic and therefore safe for consumption at the tested doses.

Medicinal components in Termitomyces mushrooms.

Termitomyces is a genus of edible mushrooms commonly consumed in Africa and Asia among the mushrooms collected from the wild. Termitomyces mushrooms grow as symbionts in the termite nests, where they produce various enzymes to help termites digest lignocellulosic substrates. Many species of Termitomyces are used by different ethnic groups with ethnomedicinal knowledge. Bioactive components that Termitomyces mushrooms contain have potential uses as antioxidants, immunomodulators, antitumors, and antimicrobials. Termitomyces also has a potential for treating neurodegenerative disorders. Here, we review the bioactive compounds from Termitomyces species that have been isolated and assayed in vitro and/or in vivo for their medicinal properties.

[Simultaneous determination of amino acids and nucleosides of three species in Termitomyces by UFLC-QTRAP-MS-MS].

A comprehensive analytical method based on UFLC-QTRAP-MS-MS was developed for the simultaneous determination of 15 kinds of amino acids and 12 kinds of nucleosides of three species in Termitomyces. The separation was carried out on a Waters XBridge Amide column (2.1 mm×100 mm,3.5 µm) with gradient elution of mobile phase of 0.2% formic acid in water-0.2% formic acid in acetonitrile at a flow rate of 0.6 mL•min⁻¹, and column temperature was maintained at 30 ℃. The target compounds were analyzed by the positive ion multiple reaction monitoring (MRM) mode. The principal component analysis(PCA) was made to standardized treatment for the comprehensive evaluation of different species in Termitomyces. The 15 kinds of amino acids and 12 kinds of nucleosides multiple constituents showed good linearity (r>0.997 3) in the range of the tested concentration.The average recoveries ranged from 95.14% to 105.0%,and the relative standard deviations were less than 5.0%. The comprehensive evaluation index obtained with PCA showed that the Termitomyces albuminosus was significantly higher than others in amino acids and in nucleosides, of which the T. aurantiacus was the best. The developed method with good repeatability and accuracy was suitable for the simultaneous determination of multiple functional substances,which provided a new basis for the comprehensive assessment and overall control of the quality of Termitomyces fungi.

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.

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.

A protein containing glucan from an edible mushroom, Termitomyces microcarpus (var).

A water-soluble protein containing glucan was isolated from an edible mushroom, Termitomyces microcarpus (var). It was found to contain 55% of carbohydrate and 40% of protein. The protein part was found to consist of glutamine, arginine, isoleucine, leucine and phenylalanine. The structure of the glucan was analyzed on the basis of total acid hydrolysis, methylation, periodate oxidation and NMR studies (1H, 13C, TOCSY, DQF-COSY, NOESY, and HMQC), and the repeating unit of the glucan is established as: --> 6)-alpha-D-Glcp-(1-->.

Analgesic and anti-inflammatory effects of the dry matter of culture broth of Termitomyces albuminosus and its extracts.

AIM OF THE STUDY: The objectives of this study were to investigate the analgesic and anti-inflammatory effects of the dry matter of culture broth (DMCB) of Termitomyces albuminosus in submerged culture and its crude saponin extract (CSE) and crude polysaccharide extract (CPE). MATERIALS AND METHODS: The analgesic effects of DMCB, CSE and CPE were evaluated with models of acetic acid-induced writhing response and formalin test in mouse. The anti-inflammatory effects of DMCB, CSE and CPE were evaluated by using models of xylene-induced mouse ear swelling and carrageen-induced mouse paw edema. RESULTS: The DMCB, CSE and CPE significantly decreased the acetic acid-induced writhing response and the licking time on the late phase in the formalin test. Treatment of DMCB (1000mg/kg), CSE (200mg/kg) or CPE (200mg/kg) inhibited the mouse ear swelling by 61.8%, 79.0% and 81.6%, respectively. In the carrageen-induced mouse paw edema test, the group treated with indomethacin showed the strongest inhibition of edema formation by 77.8% in the third hour after carrageenan administration, while DMCB (1000mg/kg), CSE (200mg/kg) and CPE (200mg/kg) showed 48.4%, 55.6% and 40.5%, respectively. CONCLUSIONS: The results suggested that DMCB of Termitomyces albuminosus possessed the analgesic and anti-inflammatory activities. Saponins and polysaccharides were proposed to be the major active constituents of Termitomyces albuminosus in submerged culture.