Selenium acumulation in submerged and industrial cultivated Coriolus versicolor mushroom

Poslednjih godina postoji veliko interesovanje za razvoj novih nutraceutika i funkcionalne hrane na bazi gljiva. Gljive poseduju svojstvo da akumuliraju teške metale iz supstrata na kome rastu. U ovoj disertaciji ispitana je sposobnost rasta i akumulacije selena iz čvrstog supstrata i tečne podloge od strane C. versicolor medicinske gljive. Kao izvori selena upotrebljeni su neorganske soli, natrijum selenat i natrijum selenit i organski izvori u obliku selenskog kvasca (Sel-Plex®-a) i selenouree. Primenjene su koncentracije od 10 mg Se L-1 i 25 mg Se L-1 za gajenje u šaržnom bioreaktoru i 10 mg Se kg-1, 25 mg Se kg-1 i 50 mg Se kg-1 računato na suvu masu supstrata za gajenje na čvrstoj podlozi. Na osnovu dobijenih rezultata pokazano je da postoji srednja, negativna, statistički značajna korelacija (p≤0.05, r=-0.54), između sadržaja neorganskog selena i prinosa plodonosnih tela i veoma jaka, negativna, statistički značajna, (p≤0.05, r = -0.89) korelacija između sadržaja organskog selena u podlozi i prinosa plodonosnih tela. Koncentracija od 10 mg Se kg-1 suvog supstrata nije statistički značajno (p≤0.05) uticala na prinos gljive u poređenju sa kontrolnim uzorkom, bez obzira na izvor selena za obogaćenje supstrata. Međutim, dodatkom 50 mg Se kg-1 suvog supstrata u obliku Sel-Plex®-a, fruktifikacija je potpuno izostala, a primenom iste koncentracije selena dodatkom selenouree su dobijena plodonosna tela izmenjene morfologije. Optimalno vreme gajenja gljive od osam dana ustanovljeno je merenjem potrošnje glukoze (HPLC metodom) tokom rasta u tečnoj podlozi. Gljiva je rasla u obliku sferičnih peleta kada je tečna podloga obogaćena selenoureom. Gljiva je formirala ugruške dispergovanih filamenata kada je tečnoj podlozi dodat Sel-Plex®, rast gljive je bio potpuno raspršen kada je upotrebljen natrijum selenit, a oba oblika rasta otežavaju separaciju dobijene biomase od podloge. Analiza skenirajućom elektronskom mikroskopijom nije potvrdila prisustvo egzopolisaharida na površini micelijuma...In recent years there has been great interest for the development of new nutraceuticals and functional foods based on mushrooms. Mushrooms are able to accumulate heavy metals from the growing substrate. In this dissertation, the ability of growth and accumulation of selenium from the solid substrate and liquid medium by C. versicolor medicinal mushrooms was analysed. As selenium sources inorganic salts such as sodium selenate and sodium selenite and organic sources in the form of selenium yeast Sel-Plex® and selenourea were used. Concentration that were applied were 10 mg Se L-1 and 25 mg Se L-1 for the cultivation in a batch bioreactor, and 10 mg Se kg-1, 25 mg Se kg-1 and 50 mg Se kg-1 based on the dry weight of the solid substrate for solid state cultivation. According to obtained results, there is a negative, medium and statistically significant correlation, (p≤0.05, r= -0.54) between the content of inorganic selenium and fruit body yield, and very strong, negative and statistically significant (p≤0.05, r = -0.89) correlation between the content of organic selenium in the growing substrate and biomass yield of the fruit body. Selenium concentration of 10 mg Se kg-1 of dry substrate did not statistically significantly (p≤0.05) affected mushroom yield compared to the control sample, regardless of the selenium source used for enrichment. However, after addition of 50 mg Se kg-1 in form of Sel-Plex®, there was no fructification at all, while the addition of the same selenium concentration using selenourea obtained fruiting bodies had altered morphology. The optimal eight day period for the submerged mushroom cultivation was established by measuring the glucose consumption (HPLC method) during mushroom growing in liquid culture. Mushrooms growing in a liquid medium enriched with selenourea formed spherical pellets. Mushroom formed clumps of dispersed filaments when Sel-Plex® was added to the liquid medium, growth was fully dispersed when sodium selenite was used, and both forms impeded separation of the biomass from the liquid medium..

The fungistatic activity of organic selenium and its application to the production of cultivated mushrooms agaricus bisporus and pleurotus spp.

The activity of organic selenium against pathogenic molds and its use as a potential selenium source in the production of enriched mushrooms were examined. The effect of commercial selenized yeast on mycelia growth was examined using a method with mycelia disks and a well diffusion method. For mushroom enrichment, different concentrations of selenium were added to a growth substrate. The results presented in this paper suggest that the most suitable concentration of selenized yeast that inhibits the growth of the mycopathogenic molds is 70-100 mg/kg of selenium. With the addition of this concentration to the substrate, mushroom fruit bodies will uptake a high level of selenium, about 100 mu g/g for Pleurotus spp., and 200 mu/g for Agaricus bisporus in dry weight of the mushroom. Thereby a double effect in the cultivation of mushrooms is achieved

The Antibacterial Activity of Coriolus versicolor Methanol Extract and Its Effect on Ultrastructural Changes of Staphylococcus aureus and Salmonella Enteritidis

The antibacterial activity of methanol extract obtained from fruiting body of industrially grown basidiomycete Coriolus versicolor was examined. The Minimum Inhibitory Concentration (MIC) values against various bacteria ranged from 0.625 to 20 mg mL(-1). versicolor expressed bactericidal activity against both Gram-positive and Gram-negative bacteria. The growth curves of Staphylococcus aureus and Salmonella enterica serovar Enteritidis, measured at 630 nm, and confirmed with macrodilution method showed that the obtained extract could inhibit the growth of tested bacteria. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), and the loss of 260-nm-absorbing material were used to examine the ultrastructural changes in bacteria induced by the extract. When S. aureus was exposed to the MIC of C. versicolor, elongated and malformed cells were observed by SEM, while S. Enteritidis treated cells appeared shorter and aggregated with ruptured cell walls. TEM revealed the formation of non-membrane-enclosed bodies and depleted inner content of S. aureus. Larger and irregular periplasmic space and deformed and scattered components of the cell envelope were observed in treated S. Enteritidis. The loss of 260-nm-absorbing material indicated that the disruptive action of the extract on cytoplasmic membrane was more pronounced in S. aureus than in S. Enteritidis treated cells. The UV and FTIR spectrophotometric analyses revealed diverse composition of C. versicolor extract and high content of total phenolics. Altogether, mushroom extracts could be used to develop nutraceuticals or drugs effective against pathogenic microorganisms

Selenium acumulation in submerged and industrial cultivated Coriolus versicolor mushroom

Poslednjih godina postoji veliko interesovanje za razvoj novih nutraceutika i funkcionalne hrane na bazi gljiva. Gljive poseduju svojstvo da akumuliraju teške metale iz supstrata na kome rastu. U ovoj disertaciji ispitana je sposobnost rasta i akumulacije selena iz čvrstog supstrata i tečne podloge od strane C. versicolor medicinske gljive. Kao izvori selena upotrebljeni su neorganske soli, natrijum selenat i natrijum selenit i organski izvori u obliku selenskog kvasca (Sel-Plex®-a) i selenouree. Primenjene su koncentracije od 10 mg Se L-1 i 25 mg Se L-1 za gajenje u šaržnom bioreaktoru i 10 mg Se kg-1, 25 mg Se kg-1 i 50 mg Se kg-1 računato na suvu masu supstrata za gajenje na čvrstoj podlozi. Na osnovu dobijenih rezultata pokazano je da postoji srednja, negativna, statistički značajna korelacija (p≤0.05, r=-0.54), između sadržaja neorganskog selena i prinosa plodonosnih tela i veoma jaka, negativna, statistički značajna, (p≤0.05, r = -0.89) korelacija između sadržaja organskog selena u podlozi i prinosa plodonosnih tela. Koncentracija od 10 mg Se kg-1 suvog supstrata nije statistički značajno (p≤0.05) uticala na prinos gljive u poređenju sa kontrolnim uzorkom, bez obzira na izvor selena za obogaćenje supstrata. Međutim, dodatkom 50 mg Se kg-1 suvog supstrata u obliku Sel-Plex®-a, fruktifikacija je potpuno izostala, a primenom iste koncentracije selena dodatkom selenouree su dobijena plodonosna tela izmenjene morfologije. Optimalno vreme gajenja gljive od osam dana ustanovljeno je merenjem potrošnje glukoze (HPLC metodom) tokom rasta u tečnoj podlozi. Gljiva je rasla u obliku sferičnih peleta kada je tečna podloga obogaćena selenoureom. Gljiva je formirala ugruške dispergovanih filamenata kada je tečnoj podlozi dodat Sel-Plex®, rast gljive je bio potpuno raspršen kada je upotrebljen natrijum selenit, a oba oblika rasta otežavaju separaciju dobijene biomase od podloge. Analiza skenirajućom elektronskom mikroskopijom nije potvrdila prisustvo egzopolisaharida na površini micelijuma...In recent years there has been great interest for the development of new nutraceuticals and functional foods based on mushrooms. Mushrooms are able to accumulate heavy metals from the growing substrate. In this dissertation, the ability of growth and accumulation of selenium from the solid substrate and liquid medium by C. versicolor medicinal mushrooms was analysed. As selenium sources inorganic salts such as sodium selenate and sodium selenite and organic sources in the form of selenium yeast Sel-Plex® and selenourea were used. Concentration that were applied were 10 mg Se L-1 and 25 mg Se L-1 for the cultivation in a batch bioreactor, and 10 mg Se kg-1, 25 mg Se kg-1 and 50 mg Se kg-1 based on the dry weight of the solid substrate for solid state cultivation. According to obtained results, there is a negative, medium and statistically significant correlation, (p≤0.05, r= -0.54) between the content of inorganic selenium and fruit body yield, and very strong, negative and statistically significant (p≤0.05, r = -0.89) correlation between the content of organic selenium in the growing substrate and biomass yield of the fruit body. Selenium concentration of 10 mg Se kg-1 of dry substrate did not statistically significantly (p≤0.05) affected mushroom yield compared to the control sample, regardless of the selenium source used for enrichment. However, after addition of 50 mg Se kg-1 in form of Sel-Plex®, there was no fructification at all, while the addition of the same selenium concentration using selenourea obtained fruiting bodies had altered morphology. The optimal eight day period for the submerged mushroom cultivation was established by measuring the glucose consumption (HPLC method) during mushroom growing in liquid culture. Mushrooms growing in a liquid medium enriched with selenourea formed spherical pellets. Mushroom formed clumps of dispersed filaments when Sel-Plex® was added to the liquid medium, growth was fully dispersed when sodium selenite was used, and both forms impeded separation of the biomass from the liquid medium..

The fungistatic activity of organic selenium and its application to the production of cultivated mushrooms Agaricus bisporus and Pleurotus spp.

The activity of organic selenium against pathogenic molds and its use as a potential selenium source in the production of enriched mushrooms were examined. The effect of commercial selenized yeast on mycelia growth was examined using a method with mycelia disks and a well diffusion method. For mushroom enrichment, different concentrations of selenium were added to a growth substrate. The results presented in this paper suggest that the most suitable concentration of selenized yeast that inhibits the growth of the mycopathogenic molds is 70-100 mg/kg of selenium. With the addition of this concentration to the substrate, mushroom fruit bodies will uptake a high level of selenium, about 100 μg/g for Pleurotus spp., and 200 μg/g for Agaricus bisporus in dry weight of the mushroom. Thereby a double effect in the cultivation of mushrooms is achieved

Morpho-Physiological and Hormonal Response of Winter Wheat Varieties to Drought Stress at Stem Elongation and Anthesis Stages

Drought stress can significantly reduce wheat growth and development as well as grain yield. This study investigated morpho-physiological and hormonal (abscisic (ABA) and salicylic (SA) acids) responses of six winter wheat varieties during stem elongation and anthesis stage as well grain yield-related traits were measured after harvest. To examine drought response, plants were exposed to moderate non-lethal drought stress by withholding watering for 45 and 65% of the volumetric soil moisture content (VSMC) for 14 days at separate experiments for each of those two growth stages. During the stem elongation phase, ABA was increased, confirming the stress status of plants, and SA showed a tendency to increase, suggesting their role as stress hormones in the regulation of stress response, such as the increase in the number of leaves and tillers in drought stress conditions, and further keeping turgor pressure and osmotic adjustment in leaves. At the anthesis stage, heavier drought stress resulted in ABA accumulation in flag leaves that generated an integrated response of maturation, where ABA was not positively correlated with any of investigated traits. After harvest, the variety Bubnjar, followed by Pepeljuga and Anđelka, did not significantly decrease the number of grains per ear and 1000 kernel weight (except Anđelka) in drought treatments, thus, declaring them more tolerant to drought. On the other hand, Rujana, Fifi, and particularly Silvija experienced the highest reduction in grain yield-related traits, considering them drought-sensitive varieties

Advances in in batch culture fermented Coriolus versicolor medicinal mushroom for the production of antibacterial compounds

Bioactive compounds obtained from Coriolus versicolor (Trametes versicolor (L: Fr) Lloyd, 1920.) mushrooms cultivated in a stirred-tank bioreactor were tested to determine their antimicrobial potential. Extracellular polysaccharides were isolated from the fermentation broth by ethanol precipitation. A methanol extract was prepared from mycelium. The cultivation conditions applied during the fermentation process provided for significant biomass 6.63 +/- 0.31 g dry weight L-1 and yield of extracellular polysaccharides (EPS) (0.74 +/- 0.12 g L-1). Microscopic analyses revealed that the mycelium grew predominately in the form of fluffy pellets. The methanol extract demonstrated very good activity against all the tested Gram-positive bacteria. Bacillus spizizeni and Staphylococcus epidermidis were the most sensitive strains (minimum inhibitory concentration (MIC) lt 0.3125 mg mL(-1)). Among the Gram-negative bacteria, Yersinia enterocolitica had the lowest MIC value, 5 mg mL(-1). Microbicidal activity of mycelia methanol extract was established in seven out of ten tested Gram-negative bacteria strains with minimum bactericidal concentration (MBC) values ranged from 20 to 40 mg mL(-1). Enterococcus faecalis and Staphylococcus aureus showed higher sensitivity to the extracellular polysaccharides (MIC values 2.5 mg mL(-1)). FTIR analysis revealed a more complex chemical composition of the methanol extract compared to EPS, which might explain the better antibacterial activity of the methanol extract. Our results suggested that the submerged cultivation of Coriolus versicolor followed by ethanol precipitation of EPS and the methanol extraction of mycelia can be a promising process to obtain biological active compounds with significant antimicrobial activity. Industrial Relevance: Mushrooms contain a large number of chemicals with potential use as antimicrobial compounds. One of the biggest challenges for providing biologically active compounds from mushrooms is short-term process standardization with a low risk for contamination. Submerged culture cultivation is the best choice for providing antimicrobial compounds from mushrooms. The submerged culture method represents an effective and energy-efficient means to produce novel antibacterial compound from mushrooms. Antibacterial activity testing revealed that methanol extract and isolated exopolysaccharides exhibited strong antibacterial activity, especially against Gram-positive bacteria