Anticancer and Immunopotentiating Activities of Crude Polysaccharides from Pleurotus nebrodensis on Mouse Sarcoma 180

Pleurotus nebrodensis is an edible and commercially available mushroom in Korea. This study was conducted in order to evaluate the anticancer and immunopotentiating activities of crude polysaccharides, extracted in methanol, neutral saline, and hot water (hereafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of P. nebrodensis. beta-Glucan and protein contents in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. nebrodensis ranged from 23.79~36.63 g/100 g and 4.45~6.12 g/100 g, respectively. Crude polysaccharides were not cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at a range of 10~2,000 microg/mL. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 11.76~27.06% in mice previously inoculated with sarcoma 180. Treatment with Fr. NaCl resulted in an increase in the numbers of spleen cells by 1.49 fold at the concentration of 50 microg/mL, compared with control. Fr. HW improved the immuno-potentiating activity of B lymphocytes through an increase in alkaline phosphatase activity by 1.65 fold, compared with control at 200 microg/mL. Maximum production of nitric oxide (14.3 microM) was recorded in the Fr. NaCl fraction at 200 microg/mL. Production of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and interleukin-6 (IL-6) was significantly higher, compared to control, and IL-6 production was highest, in contrast to TNF-alpha, IL-1beta, and positive control, concanavalin at the tested concentration of the various fractions. Results of the current study suggest that polysaccharides extracted from P. nebrodensis have a strong anticancer effect and may be useful as an ingredient of biopharmaceutical products for treatment of cancer.

Immunomodulating and Antitumor Activities of Panellus serotinus Polysaccharides

This study was initiated in order to investigate the anticancer and immunomodulating activities of crude polysaccharides extracted in methanol, neutral saline, and hot water (hereinafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of Panellus serotinus. Content of beta-glucan and protein in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. serotinus ranged from 22.92~28.52 g/100 g and 3.24~3.68 g/100 g, respectively. In vitro cytotoxicity tests, none of the various fractions of crude polysaccharides were cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at the tested concentration. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 23.53~44.71% in mice previously inoculated with sarcoma 180. Treatment with Fr. HW resulted in an increase in the numbers of spleen cells by 1.3 fold at the concentration of 50 microg/mL compared with control. Treatment with Fr. NaCl resulted in improvement of the immuno-potentiating activity of B lymphocytes by increasing the alkaline phosphatase activity by 1.4 fold, compared with control, at the concentration of 200 microg/mL. Among the three fractions, maximum nitric oxide (13.48 microM) was recorded at 500 microg/mL in Fr. HW. Production of tumor necrosis factor alpha, interleukin-1beta, and interleukin-6 was significantly higher, compared to the positive control, concanavalin A, at the tested concentration. Therefore, treatment with crude polysaccharides extracted from the fruiting body of P. serotinus could result in improvement of antitumor activity.

Antihyperlipidemic Effect of Dietary Lentinus edodes on Plasma, Feces and Hepatic Tissues in Hypercholesterolemic Rats

We investigated diet supplementation with shiitake mushroom fruiting bodies on biochemical and histological changes in hypercholesterolemic rats. Six-wk old female Sprague-Dawley albino rats were divided into three groups of 10 rats each. A diet containing 5% Lentinus edodes fruiting bodies given to hypercholesterolemic rats reduced plasma total cholesterol, triglyceride, low-density lipoprotein (LDL), total lipid, phospholipids, and the LDL/high-density lipoprotein ratio by 34.33, 53.21, 75.00, 34.66, 25.73, and 71.43%, respectively. Feeding mushroom also significantly reduced body weight in hypercholesterolemic rats. However, it had no detrimental effects on plasma albumin, total bilirubin, direct bilirubin, creatinine, blood urea nitrogen, uric acid, glucose, total protein, calcium, sodium, potassium, chloride, inorganic phosphate, magnesium, or enzyme profiles. Feeding mushroom increased total lipid and cholesterol excretion in feces. The plasma lipoprotein fraction, separated by agarose gel electrophoresis, indicated that L. edodes significantly reduced plasma beta and pre-beta-lipoprotein but increased alpha-lipoprotein. A histological study of hepatic cells by conventional hematoxylin-eosin and oil red-O staining showed normal findings for mushroom-fed hypercholesterolemic rats. These results suggest that shiitake mushrooms could be recommended as a natural cholesterol lowering substance in the diet.

Assessment of Antioxidant and Phenolic Compound Concentrations as well as Xanthine Oxidase and Tyrosinase Inhibitory Properties of Different Extracts of Pleurotus citrinopileatus Fruiting Bodies

Cellular damage caused by reactive oxygen species has been implicated in several diseases, thus establishing a significant role for antioxidants in maintaining human health. Acetone, methanol, and hot water extracts of Pleurotus citrinopileatus were evaluated for their antioxidant activities against beta-carotene-linoleic acid and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, reducing power, ferrous ion-chelating abilities, and xanthine oxidase inhibitory activities. In addition, the tyrosinase inhibitory effects and phenolic compound contents of the extracts were also analyzed. Methanol and acetone extracts of P. citrinopileatus showed stronger inhibition of beta-carotene-linoleic acid compared to the hot water extract. Methanol extract (8 mg/mL) showed a significantly high reducing power of 2.92 compared to the other extracts. The hot water extract was more effective than the acetone and methanole extracts for scavenging DPPH radicals. The strongest chelating effect (92.72%) was obtained with 1.0 mg/mL of acetone extract. High performance liquid chromatography analysis detected eight phenolic compounds, including gallic acid, protocatechuic acid, chlorogenic acid, ferulic acid, naringenin, hesperetin, formononetin, and biochanin-A, in an acetonitrile and hydrochloric acid (5 : 1) solvent extract. Xanthine oxidase and tyrosinase inhibitory activities of the acetone, methanol, and hot water extracts increased with increasing concentration. This study suggests that fruiting bodies of P. citrinopileatus can potentially be used as a readily accessible source of natural antioxidants.

Cultural Conditions for Mycelial Growth and Molecular Phylogenetic Relationship in Different Wild Strains of Schizophyllum commune

The common split-gilled mushroom, Schizophyllum commune is found throughout the world on woody plants. This study was initiated to evaluate conditions for favorable vegetative growth and to determine molecular phylogenetic relationship in twelve different strains of S. commune. A suitable temperature for mycelial growth was obtained at 30degrees C. This mushroom grew well in acidic conditions and pH 5 was the most favorable. Hamada, glucose peptone, Hennerberg, potato dextrose agar and yeast malt extract were favorable media for growing mycelia, while Lilly and glucose tryptone were unfavorable. Dextrin was the best and lactose was the less effective carbon source. The most suitable nitrogen sources were calcium nitrate, glycine, and potassium nitrate, whereas ammonium phosphate and histidine were the least effective for the mycelial growth of S. commune. The genetic diversity of each strain was investigated in order to identify them. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 129 to 143 bp and 241 to 243 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical. A phylogenetic tree of the ITS region sequences indicated that the selected strains were classified into three clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) with 20 arbitrary primers. Twelve primers efficiently amplified the genomic DNA. The number of amplified bands varied depending on the primers used or the strains tested. The average number of polymorphic bands observed per primer was 4.5. The size of polymorphic fragments was obtained in the range of 0.2 to 2.3 kb. These results indicate that the RAPD technique is well suited for detecting the genetic diversity in the S. commune strains tested.

Mycelial Propagation and Molecular Phylogenetic Relationships of Commercially Cultivated Agrocybe cylindracea based on ITS Sequences and RAPD

This study evaluated the optimal vegetative growth conditions and molecular phylogenetic relationships of eleven strains of Agrocybe cylindracea collected from different ecological regions of Korea, China and Taiwan. The optimal temperature and pH for mycelial growth were observed at 25degrees C and 6. Potato dextrose agar and Hennerberg were the favorable media for vegetative growth, whereas glucose tryptone was unfavorable. Dextrin, maltose, and fructose were the most effective carbon sources. The most suitable nitrogen sources were arginine and glycine, whereas methionine, alanine, histidine, and urea were least effective for the mycelial propagation of A. cylindracea. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The sequence of ITS2 was more variable than that of ITS1, while the 5.8S sequences were identical. The reciprocal homologies of the ITS sequences ranged from 98 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) using 20 arbitrary primers. Fifteen primers efficiently amplified the genomic DNA. The average number of polymorphic bands observed per primer was 3.8. The numbers of amplified bands varied based on the primers and strains, with polymorphic fragments ranging from 0.1 to 2.9 kb. The results of RAPD analysis were similar to the ITS region sequences. The results revealed that RAPD and ITS techniques were well suited for detecting the genetic diversity of all A. cylindracea strains tested.

Phylogenetic Relationship in Different Commercial Strains of Pleurotus nebrodensis Based on ITS Sequence and RAPD

The molecular phylogeny in nine different commercial cultivated strains of Pleurotus nebrodensis was studied based on their internal transcribed spacer (ITS) region and RAPD. In the sequence of ITS region of selected strains, it was revealed that the total length ranged from 592 to 614 bp. The size of ITS1 and ITS2 regions varied among the strains from 219 to 228 bp and 211 to 229 bp, respectively. The sequence of ITS2 was more variable than ITS1 and the region of 5.8S sequences were identical. Phylogenetic tree of the ITS region sequences indicated that selected strains were classified into five clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by RAPD with 20 arbitrary primers. Twelve primers were efficient to applying amplification of the genomic DNA. The sizes of the polymorphic fragments obtained were in the range of 200 to 2000 bp. RAPD and ITS analysis techniques were able to detect genetic variation among the tested strains. Experimental results suggested that IUM-1381, IUM-3914, IUM-1495 and AY-581431 strains were genetically very similar. Therefore, all IUM and NCBI gene bank strains of P. nebrodensis were genetically same with some variations.

Vegetative Growth and Phylogenetic Relationship of Commercially Cultivated Strains of Pleurotus eryngii based on ITS sequence and RAPD

Pleurotus eryngii, known as king oyster mushroom has been widely used for nutritional and medicinal purposes. This study was initiated to screen the suitable conditions for mycelial growth and to determine the phylogenetic relationship of the selected strains. Optimal mycelial growth was observed at 30degrees C and minimum mycelial growth observed at 10degrees C. This mushroom tolerates a broad pH range for mycelial growth, with most favorable growth observed at pH 6. Results also indicated that glucose peptone, yeast malt extract and mushroom complete media were favorable growth media, while Hennerberg and Hoppkins media were unfavorable. Dextrin was the best and xylose the least effective carbon sources. Results revealed that inorganic nitrogen sources were less effective than organic sources for the mycelial growth of P. eryngii. Investigation of genetic diversity is necessary to identify the strains. The ITS region of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 214 to 222 bp and 145 to 236 bp, respectively. The sequence of ITS2 was more variable than that of ITS1, and the 5.8S sequences were identical. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into six clusters. Fourteen IUM and ATCC-90212 strains were also analyzed by RAPD with 20 arbitrary primers. Fourteen of these primers were efficiently amplified the genomic DNA. The number of amplified bands varied with the primers and strains, with polymorphic fragments in the range from 0.2 to 2.3 kb.

Comparative Effects of Oyster Mushrooms on Lipid Profile, Liver and Kidney Function in Hypercholesterolemic Rats

Comparative effects of oyster mushrooms on plasma and fecal lipid profiles and on liver and kidney function were evaluated in hyper and normocholesterolemic rats. Feeding of hypercholesterolemic rats a 5% powder of oyster mushrooms (Pleurotus ostreatus, P. sajor-caju and P. florida) reduced the plasma total cholesterol level by 37%, 21% and 16%, respectively and reduced the triglyceride level by 45%, 24% and 14%, respectively. LDL/HDL ratio decreased by 64%, 45% and 41% for P. sajor-caju, P. ostreatus and P. florida fed rats, respectively. Mushroom feeding also reduced body weight in hypercholesterolemic rats. However, it had no adverse effect on plasma bilirubin, creatinin and urea nitrogen level. Mushroom feeding also increased the total lipid and cholesterol excretion in the feces. The present study reveals that feeding of 5% oyster mushroom powder does not have detrimental effects on the liver and kidneys rather may provide health benefits for the cardiovascular-related complication by decreasing the atherogenic lipid profiles.

Vegetative Growth of Four Strains of Hericium erinaceus Collected from Different Habitats

Vegetative growth of four different strains of Hericium erinaceus was observed. The temperature suitable for optimal mycelial growth was determined to be 25degrees C, with growth observed in the extend temperature range of 20~30degrees C. The different strains of this mushroom showed distinct pH requirements for their optimum vegetative growth, with the most favorable growth observed at pH 6. Considering vegetative mycelial growth, PDA, YM, Hennerberg, Hamada, and Glucose peptone were the most favorable media, and Czapek Dox, Hoppkins, Glucose tryptone, and Lilly were the most unfavorable media for these mushroom strains. With the exception of lactose, most of the carbon sources assayed demonstrated favorable vegetative growth of H. erinaceus. For mycelial growth, the most suitable nitrogen source was alanine and the most unsuitable was histidine. Oak sawdust medium supplemented with 10~20% rice bran was the best for mycelial growth of the mushroom.

Nutritional Analysis of Cultivated Mushrooms in Bangladesh - Pleurotus ostreatus, Pleurotus sajor-caju, Pleurotus florida and Calocybe indica

Mushroom cultivation has been started recently in Bangladesh. Awareness of the nutritional and medicinal importance of mushrooms is not extensive. In this study, the nutritional values of dietary mushrooms- Pleurotus ostreatus, Pleurotus sajorcaju, Pleurotus florida and Calocybe indica that are very popular among the cultivated mushrooms in Bangladesh have been determined. These mushrooms were rich in proteins (20~25%) and fibers (13~24% in dry samples) and contained a lower amount of lipid (4 to 5%). The carbohydrate contents ranged from 37 to 48% (on the basis of dry weight). These were also rich in mineral contents (total ash content is 8~13%). The pileus and gills were protein and lipid rich and stripe was carbohydrate and fiber-rich. The moisture content of mushrooms ranged from 86 to 87.5%. Data of this study suggest that mushrooms are rich in nutritional value.

Physicochemical Requirement for the Vegetative Growth of Schizophyllum commune Collected from Different Ecological Origins

Schizophyllum commune is an edible and medicinal mushroom widely distributed in the world. The optimal growth conditions for the mycelia of 10 strains of the fungus were investigated. The temperature suitable for the mycelial growth and density was obtained at 30~35degrees C. Among the tested conditions, the minimum mycelial growth was found at 15degrees C. In case of pH, the most favorable growth was found at pH 5. The results indicated that this mushroom well adapted to high temperature and low pH for its mycelial growth. Considering growth phenotype of mycelia, Hamada, Hennerberg, PDA and YM were the most suitable and Lilly, Glucose triptone, Glucose peptone and Hoppkins were the most unfavorable among tested media for the mycelial growth of S. commune. Out of tested carbon sources, dextrin and fructose were the most suitable and lactose, mannose and sorbitol were the unsuitable for the fungus. Compact mycelial density was obtained from most of the carbon sources. Among used nitrogen sources, calcium nitrate, potassium nitrate and alanine were the most appropriate and the most incompatible were ammonium phosphate, histidine, urea and arginine for mycelial growth of S. commune on the culture media. Calcium nitrate, histidine and potassium nitrate showed moderately thin or thin, and rest of nitrogen sources showed compact or moderately compact mycelial density.

The Artificial Cultivation of Oudemansiella mucida on the Oak Sawdust Medium

To produce fruiting bodies of Oudemansiella mucida, porcelain fungus, on the oak sawdust medium, additives suitable for the mycelial growth and fruiting body formation were screened. In general, the mycelial growth of the three strains of O. mucida used in this study have been good on oak sawdust mixed rice bran of 20~30%. The mycelia incubated in potato dextrose broth for 7 days were inoculated on oak sawdust medium supplemented with various ratios of rice bran and incubated for 30 days at 25degrees C in the dark condition until the mycelia of O. mucida fully colonized the media from top to bottom. Then, top surface of the media in the bottles were horizontally scratched with a spatula and filled with tap water for 3 hours. To induce the primordial formation of O. mucida, the bottles were transferred to the mushroom cultivating room under 12 hrs of light (350 lux) and dark condition with relative humidity of 95% at 17degrees C. The primordia of O. mucida were formed on the surface of oak sawdust media after 7 days of incubation. The mature fruiting bodies were observed 5 days after primordial formation. The fruiting bodies O. mucida were formed on oak sawdust medium mixed with 5 to 30% rice bran. However, abundant fruiting-bodies of O. mucida were produced in oak sawdust medium supplemented with 20% rice bran. This is the first report associated with an artificial fruiting body production of O. mucida in Korea.

The Optimal Culture Conditions for the Mycelial Growth of Oudemansiella radicata

Oudemansiella radicata, one of edible mushrooms belonging to Tricholomataceae of Basidiomycota, has been known to exhibit outstanding therapeutic effects on the hypertension caused by high blood pressure and inhibitory effects on the sarcoma 180 and Erhrlich carcinoma of mice. As one of preliminary experiments for producing fruiting-body of O. radicata, this study was carried out to obtain the basic information for culture conditions of mycelial growth of the fungus. The optimal temperature and pH for the mycelial growth were 25degrees C and pH 6, respectively. The medium for favorable mycelial growth of O. radicata was shown in the Lilly medium, whereas compact mycelial density was found in Hamada medium. The carbon and nitrogen sources promoting for mycelial growth of O. radicata were xylose and alanine, respectively. The optimum C/N ratio was about 20 : 1 in case that 3% glucose was supplimented to the basal medium as a carbon source.

The Characteristics of Cultural Conditions for the Mycelial Growth of Macrolepiota procera

Macrolepiota procera, one of edible mushrooms belongs to Agaricaceae of Basidiomycota, has a good taste and good medicinal value. As a preliminary study for the development of artificial cultivation method of edible mushroom, cultural characteristics of M. procera was investigated on various culture media under different environmental conditions. Mycelial growth was compared on culture media composed of various carbon and nitrogen sources, and C/N ratios. The optimal conditions for the mycelial growth were 30degrees C and pH 7. M. procera showed the rapid mycelial growth in the PDA media. The optimal carbon and nitrogen sources were maltose and glycine, respectively. The optimum C/N ratio was about 10 : 1 in case that 1% glucose was supplemented to the basal media as carbon source.

Culture Conditions Affecting the Optimal Mycelial Growth of Cystoderma amianthinum

Cystoderma amianthinum, one of edible fungi belongs to Agaricaceae of Basidiomycota, has a good taste and flavor. This study was carried out to obtain the basic informations for the optimum mycelial growth of C. amianthinum. The optimal conditions for the mycelial growth were 25degrees C and pH 5 in potato dextrose agar (PDA). C. amianthinum showed the favorable growth in the PDA and yeast malt extract agar (YMA). The favorable carbon and nitrogen sources promoting mycelial growth were fructose and histidine, respectively. The optimum C/N ratio was about 30 : 1 in case that 1% glucose was supplemented to the basal medium as a carbon source.

Distribution of Pathogenic Genes and Molecular Typing of Yersinia pseudotuberculosis Isolated from Spring Water in Seoul

In order to investigate pathogenic genes and genetic relationships of Y. pseudotuberculosis strains, We isolated 9 strains of Y. pseudotuberculosis from 380 spring water sites in Seoul from 2000 to 2003. All isolates were distributed to the northeast area in Seoul. The isloates were analyzed for chromosomal virulence gene (inv) and plasmid-borne genes (yadA and lcrF) using PCR to assume pathogenicity. As a result, all isolates were positive for the inv gene, but only five isolates (55.6%) were positive for the yadA and lcrF genes. RAPD and PCR-ribotyping were tested and all isolates were grouped with 90% similarity. RAPD revealed 4 clusters and PCR-ribotyping revealed 2 clusters. The result of this experiments confirmed the view that RAPD had better powerful discrimination than PCR-ribotyping and RAPD typing was effective to distinguish between various strains of Y. pseudotuberculosis from spring water.