Fumio Eguchi

Regeneration of protoplasts from hyphal strands of Volvariella volvacea

A series of experiments on the preparation and regeneration of protoplasts from hyphal strands ofVolvariella volvacea (Bull. ex. Fr.) Singer were conducted with the aim of optimizing the conditions for its efficient regeneration. One commercial (Vvcl) and two wild (EAAC-0001 and EAAC-0002) strains ofV. volvacea from the Philippines were used and subjected to varying conditions to determine the most efficient means for regeneration of their protoplasts. The effects of age and type of strain, pH, type and concentration of osmotic stabilizer, enzymatic composition, treatment time, temperature, reciprocal frequency during enzymatic lysis of the cell wall, and centrifugation conditions were investigated. Results showed that the three strains ofV. volvacea had varying responses in terms of yield, size, and ability of their protoplasts to regenerate into the protoplast regeneration medium. Among the three strains, EAAC-0002 had the highest rate of regeneration. The 5-day-old culture ofV. volvacea, when subjected to a combination of 2% Novozyme 234 and 0.2% chitinase in 0.6M mannitol (pH 6.0) for 3 h at 30°C, 90 strokes/min and centrifuged at 1100 g for 10 min; produced an efficient yield of protoplasts with a relatively high regeneration rate.

Effects of a Hot-Water Extract of Porcini (Boletus aestivalis) Mushrooms on the Blood Pressure and Heart Rate of Spontaneously Hypertensive Rats

The repeated once-daily oral administration of a hot-water extract of porcini, Boletus aestivalis, mushrooms (WEP) to spontaneously hypertensive rats (SHR) for 18 weeks decreased the systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate. The WEP administration also decreased blood urea nitrogen (BUN), creatinine (Cre), and triglyceride (TG), and increased high-density lipoprotein-cholesterol (HDL-C) in the blood, suggesting that WEP improved the status of hypertension, as well as the high heart rate and metabolic abnormalities involved in hypertension.

Effects of Psilocybe argentipes on Marble-Burying Behavior in Mice

Psilocybe argentipes is a hallucinogenic mushroom. The present study examined the effects of P. argentipes on marble-burying behavior, which is considered an animal model of obsessive-compulsive disorder. P. argentipes significantly inhibited marble-burying behavior without affecting locomotor activity as compared with the same dose of authentic psilocybin. These findings suggest that P. argentipes would be efficient in clinical obsessive-compulsive disorder therapy.

Physiological considerations for efficient mycelial colonization of Philippine strains of Volvariella volvacea

The nutritional and physical requirements for the efficient mycelial colonization ofVolvariella volvacea (Bull. ex. Fr.). Singer were elucidated with the percentage mycelial colonization and density as references. This investigation was limited to the evaluation of two commercial strains (designated Vvc1 and Vvc2) and two wild strains (designated EAAC-0001 and EAAC-0002) ofV. volvacea from the Philippines with the aim of providing baseline data on their physiological requirements. The four strains ofV. volvacea had varying preferences for carbon. Vve1 preferred polysaccharides (starch and cellulose), whereas Vvc2 grew luxuriantly at a relatively rapid rate in sugar alcohol (sorbitol). The two wild strains preferred starch as a carbon source. In terms of nitrogen utilization, soytone, peptone, and glycine supported efficient mycelial colonization of the four strains. The vitamin utilization test revealed that ascorbic acid, calcium pantothenate, and biotin are good sources. The mycelial growth performances of the strains were also evaluated on six dehydrated mycological media. Efficient colonization of Vvc1, Vvc2, and EAAC-0002 with dense mycelial growth was noted in mycological agar. EAAC-0001, on the other hand, grew more efficiently in malt extract agar. The Philippine strains ofV. volvacea grew luxuriantly when incubated at 35°C and pH 8.0 under dark and sealed conditions. Moreover, the relatively higher moisture content (70%) of the oolong tea leaf formulation favorably stimulated efficient mycelial colonization. Under optimum physiological conditions, Vvc1, Vvc2, and EAAC-0002 were fast-growing strains, whereas EAAC-0001 was a moderately growing type.

Pharmacological effects of Ganoderma lucidum collected from ume (Japanese apricot) trees

We performed functional evaluation of the fruiting bodies of Ganoderma lucidum growing on ume trees (Japanese apricot, Prunus mume), and determined the suitability of pruned ume branches as a basic component of culture medium for this mushroom. We observed that all tested functional activities of the fruiting bodies of G. lucidum collected from ume trees were higher than those collected from other broadleaf trees or cultured artificially; the functional tests were angiotensin I-converting enzyme inhibitory activity, a platelet aggregation inhibition test, and an interleukin-8 (IL-8) gene expression inhibition test. When extracts from fruiting bodies of G. lucidum were orally administered to spontaneous hypertensive rats, hypotensive effects were found. Freeze drying was the most suitable procedure for preservation of the extracts, and the activities of 30% ethanol extracts and 30% methanol extracts were higher than those of hot-water extracts. The highest functional activities for extracts from G. lucidum mycelia cultured on sawdust media were for sawdusts based on ume wood.

Improvement of Diet-induced Obesity by Ingestion of Mushroom Chitosan Prepared from Flammulina velutipes

The anti-obesity effects of mushroom chitosan prepared from Flammulina velutipes were investigated using an animal model with diet-induced obesity. In this study, 5-week-old imprinting control region (ICR) mice were divided into six groups of 10 mice each and fed different diets based on the MF powdered diet (standard diet) for 6 weeks: standard diet control group, high-fat diet control group (induced dietary obesity) consisting of the standard diet and 20% lard, and mushroom chitosan groups consisting of the high-fat diet with mushroom chitosan added at 100, 500, 1,000, and 2,000 mg/kg body weight. On the final day of the experiment, mean body weight was 39.1 g in the high-fat control group and 36.3 g in the 2,000 mg/kg mushroom chitosan group, compared to 35.8 g in the standard diet control group. In the mushroom chitosan groups, a dose-dependent suppression of weight gain and marked improvements in serum triglycerides, total cholesterol, LDL-cholesterol, and HDL-cholesterol were found. The mushroom chitosan groups showed fewer and smaller fat deposits in liver cells than the high-fat diet control group, and liver weight was significantly reduced. Glutamic oxaloacetic transaminase (GOT) and glutamate pyruvic transaminase (GPT), which are indices of the hepatic function, all showed dose-dependent improvement with mushroom chitosan administration. These results suggested that mushroom chitosan acts to suppress enlargement of the liver from fat deposition resulting from a high-fat diet and to restore hepatic function. The lipid content of feces showed a marked increase correlated with the mushroom chitosan dose. These findings suggest the potential use of mushroom chitosan as a functional food ingredient that contributes to the prevention or improvement of dietary obesity by inhibiting digestion and absorption of fats in the digestive tract and simultaneously promotes lipolysis in adipocytes.

Immunomodulatory and Anti-IBDV Activities of the Polysaccharide AEX from Coccomyxa gloeobotrydiformis

A number of polysaccharides have been reported to show immunomodulatory and antiviral activities against various animal viruses. AEX is a polysaccharide extracted from the green algae, Coccomyxa gloeobotrydiformis. The aim of this study was to examine the function of AEX in regulating the immune response in chickens and its capacity to inhibit the infectious bursal disease virus (IBDV), to gain an understanding of its immunomodulatory and antiviral ability. Here, preliminary immunological tests in vitro showed that the polysaccharide AEX can activate the chicken peripheral blood molecular cells’ (PBMCs) response by inducing the production of cytokines and NO, promote extracellular antigen presentation but negatively regulate intracellular antigen presentation in chicken splenic lymphocytes, and promote the proliferation of splenic lymphocytes and DT40 cells. An antiviral analysis showed that AEX repressed IBDV replication by the deactivation of viral particles or by interfering with adsorption in vitro and reduced the IBDV viral titer in the chicken bursa of Fabricius. Finally, in this study, when AEX was used as an adjuvant for the IBDV vaccine, specific anti-IBDV antibody (IgY, IgM, and IgA) titers were significantly decreased. These results indicate that the polysaccharide AEX may be a potential alternative approach for anti-IBDV therapy and an immunomodulator for the poultry industry. However, more experimentation is needed to find suitable conditions for it to be used as an adjuvant for the IBDV vaccine.