Characterization of vitamin B12 compounds in the fruiting bodies of shiitake mushroom (Lentinula edodes) and bed logs after fruiting of the mushroom

This study determined the vitamin B₁₂ content in commercially available dried fruiting bodies of shiitake mushroom, Lentinula edodes. The vitamin B₁₂ contents in dried donko-type fruiting bodies with closed caps (5.61 ± 3.90 μg/100 g dry weight), did not significantly differ from those of dried koushin-type fruiting bodies with open caps (4.23 ± 2.42 μg/100 g dry weight). The bed logs after fruiting of the mushroom also contained the vitamin B₁₂ levels similar to that in the dried shiitake fruiting bodies. To determine whether the dried shiitake fruiting bodies and their bed logs contained vitamin B₁₂ or other corrinoid compounds that are inactive in humans, we purified corrinoid compounds using an immunoaffinity column and identified vitamin B₁₂ using vitamin B₁₂-dependent Escherichia coli 215 bioautograms and liquid chromatography-electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) chromatograms. Dried shiitake fruiting bodies rarely contained an unnatural corrinoid vitamin B₁₂[c-lactone] that is inactive in humans. Given that shiitake mushroom lacks the ability to synthesize vitamin B₁₂ de novo, the vitamin B₁₂ found in dried shiitake fruiting bodies must have been derived from the bed logs.     

Seasonal Variation of Cadmium, Copper, and Lead Concentrations in Fish from a Freshwater Lake

This paper aims to compare the Cd, Cu, and Pb concentration in the bone, gills, and muscle of the seven fish species sampled during normal and dry seasons. Cadmium, Cu, and Pb concentrations varied significantly depending on the type of the tissue and season. Bone samples of the Osteochilus hasseltii showed the highest concentrations of Pb (6.08 μg/g dw) during September (the dry season), whereas bone samples of the Puntioplites bulu showed the lowest concentrations of Cd (0.08 μg/g dw) during September. Muscle samples of the P. bulu indicated the highest concentrations of Cu (2.58 μg/g dw) during March (the normal season). On the other hand, muscle samples of Channa straitus sowed the lowest concentrations of Cd (0.04 μg/g dw) during July (the dry season). Gills samples of the O. hasseltii showed the highest concentration of Pb (6.56 μg/g dw) during March (the normal season), while gills samples of C. straitus indicated the lowest concentration of Cd (0.06 μg/g dw) during July (the dry season).     

Comparison of plant-based expression platforms for the heterologous production of geraniol

We compared the ability of different plant-based expression platforms to produce geraniol, a key metabolite in the monoterpenoid branch of the terpenoid indole alkaloid biosynthesis pathway. A geraniol synthase gene isolated from Valeriana officinalis (VoGES) was stably expressed in different tobacco systems. Intact plants were grown in vitro and in the greenhouse and were used to generate cell suspension and hairy root cultures. VoGES was also transiently expressed in N. benthamiana. The highest geraniol content was produced by intact transgenic plants grown in vitro (48 μg/g fresh weight, fw), followed by the transient expression system (27 μg/g fw), transgenic plants under hydroponic conditions in the greenhouse and cell suspension cultures (16 μg/g fw), and finally hairy root cultures (9 μg/g fw). Differences in biomass production and the duration of cultivation resulted in a spectrum of geraniol productivities. Cell suspension cultures achieved a geraniol production rate of 1.8 μg/g fresh biomass per day, whereas transient expression produced 5.9 μg/g fresh biomass per day (if cultivation prior to agroinfiltration is ignored) or 0.5 μg/g fresh biomass per day (if cultivation prior to agroinfiltration is included). The superior productivity, strict process control and simple handling procedures available for transgenic cell suspension cultures suggest that cells are the most promising system for further optimization and ultimately for the scaled-up production of geraniol.     

Successful large-scale production of fruiting bodies of Laetiporus sulphureus (Bull.: Fr.) Murrill on an artificial substrate

Laetiporus sulphureus is an edible wood-rotting basidiomycete fungus whose fruiting bodies contain substances with verified therapeutic evidences and large amounts of α-(1 → 3)-glucan which is used as an effective inducer of microbial α-(1 → 3)-glucanases. However, production of mature fruiting bodies of this species under artificially controlled conditions has not been reported until now. Here, we provide the first report of successful initiation and development of L. sulphureus fruiting bodies in large-scale experiments. Twelve Laetiporus strains were isolated from a natural habitat. A synthetic log production system with a substrate composed of a mixture of sawdust enriched with organic and inorganic additives was developed. It was found that shocking the fungus mycelium with cold water or low temperature was the only suitable method for forced fruiting of L. sulphureus strains. Primordia of two strains were initiated already after 5–6 days from induction, and after another 2 days, they began to develop into fruiting bodies. Carpophores appeared fastest on substrates with high organic supplementation (40–45 %) and a low moisture content (40 %). The resulting mature fruiting bodies reached a weight of 200–300 g. The method of cultivation presented in this paper opens the way to commercial production of this valuable basidiomycete.     

Arsenic and Arsenic Species in Cultured Oyster (Crassostrea gigas and C. corteziensis) from Coastal Lagoons of the SE Gulf of California, Mexico

The aim of this study was to evaluate the bioavailability of arsenic (As) through cultured oyster Crassostrea gigas and Crassostrea corteziensis from four coastal lagoons (SE Gulf of California). Organisms were collected in two seasons (rainy and dry season), and they were analyzed for total arsenic and chemical speciation of this element. The concentrations of As in oyster soft tissue fluctuated between 5.44 and 9.56 μg/g for rainy season and 6.46 and 8.33 μg/g for dry season (dry weight) in C. gigas. In C. corteziensis, the As concentrations were <5 μg/g for both seasons (dry weight). Arsenic speciation indicated arsenobetaine as the major arseno-compound accounting for 43.2–76.3 % of total content of As. Lower contributions were obtained for non-extractable As (11.3–17.5 %) and other molecules such as arsenocholine and methyl-arsonate (<5 %). Inorganic arsenic was detectable in only two samples, at concentrations lower than <0.1 μg/g. These As data are the first generated for these mollusks in NW Mexico and indicate that C. gigas and C. corteziensis farmed in this area are safe for human consumption in terms of arseno-compounds.     

Mineral composition variation in the shells of freshwater mussel Anodonta woodiana at different growth stages

The levels of mineral element Na, Mg, K, Ca, Cr, Mn, Fe, Co, Ni, Cu, Zn, Se, Mo, Al, As, Ag, Cd, and Tl were quantified in the whole shells of the freshwater bivalve Anodonta woodiana at three different growth stages (i.e. J1 juveniles of 1 month old, J2 juveniles of 3.5 months old, and adults of 36 months old). The concentrations of Na and Al were different between different growth stages (p < 0.05). The highest Na concentrations (2715 ± 86 μg/g dry weight) were found in J2 juveniles. The highest Al concentrations (303.9 ± 5.95 μg/g dry weight) were found in J1 juveniles. Manganese concentrations (517.0 ± 47.98 μg/g dry weight) were significantly higher in J2 juveniles than in J1 juveniles (432.3 ± 9.87 μg/g dry weight) (p < 0.05). Copper concentrations (27.32 ± 0.15 μg/g dry weight) were significantly higher in J1 juveniles than in J2 juveniles (26.21 ± 0.86 μg/g dry weight) and adults (24.74 ± 1.43 μg/g dry weight) (p < 0.05). Burdens of Na, Ca, Mn, Fe, Co, Cu, Mo, Ag, and Tl were positively correlated with the shell length (p < 0.05). These findings can possibly contribute to an understanding of elemental requirements for shell growth and, hence, facilitate improvement of survival and growth rates during artificial mussel culture.     

Bioaccumulation of Trace Mercury in Trophic Levels of Benthic, Benthopelagic, Pelagic Fish Species, and Sea Birds from Arvand River, Iran

In this study, concentration of mercury was determined in the trophic levels of benthic, benthopelagic, pelagic fish species, and river birds from Arvand River, located in the Khuzestan province in the lowlands of southwestern Iran at the head of the Persian Gulf. The order of mercury concentrations in tissues of the fish species was as follows: liver>gill>muscle and in tissues of the kingfisher species was as follows: feather>liver>kidney>muscle. Therefore, liver in fish and feather in kingfisher exhibited higher mercury concentration than the other tissues. There was a positive correlation between mercury concentrations in fish and kingfisher species with size of its food items. We expected to see higher mercury levels in tissues of female species because they are larger and can eat larger food items. The results of this study show that the highest mean mercury level were found in the kingfisher (Anas crecca), followed by benthic (Epinephelus diacanthus), benthopelagic (Chanos chanos), and pelagic fish (Strongylura strongylura). Mean value of mercury in fish species, S. strongylura were (0.61 μg g^?1 dry weight), C. chanos (0.45 μg g^?1 dry weight), E. diacanthus (0.87 μg g^?1 dry weight), and in kingfisher species A. crecca was (2.64 μg g^?1 dry weight). Significant correlation between mercury concentration in fish and kingfisher may be related to high variability of mercury in the fish.     

Fusarium fungi and associated metabolites presence on grapes from Slovakia

Toxinogenic Fusarium species were identified on grape berries from Slovak vineyards, and their toxic metabolites were analysed by HPLC-MS/MS. F. subglutinans, F. oxysporum, F. proliferatum, F. semitectum, F. solani, F. subglutinans, and F. verticillioides were found with varying frequency. F. oxysporum and F. proliferatum, cultured in vitro on Czapek yeast autolysate agar and yeast extract sucrose agar, produced beauvericin, in the range from 3,265 to 13,400 μg/kg, and fusaproliferin in high concentration, ranging from 49,850 to 259,500 μg/kg. A maximum value of 2.24 μg/kg has been observed for beauvericin in dried grape berries. Fumonisin B_1, and fumonisin B₂ were also identified, and the observed levels ranged from 500 to 2,040 μg/kg. Over 2 years (namely 2008 and 2009) many other metabolites have been identified and analysed in grape berries, in particular: avenacein Y, apicidin, aurofusarin, chlamydosporol, 2-amino-14,16-dimethyloctadecan-3-ol, enniatin A, enniatin A_1, enniatin B_2, enniatin B_3, and equisetin.     

Nanofiltration of polysaccharides from Agaricus subrufescens

A simplified submerged airlift cultivation was established for the production of biomass from Agaricus subrufescens. In this work, soluble polysaccharides extracted from fungal mycelium, fruiting bodies, and the residual culture media were concentrated by nanofiltration. Total and high molar mass polysaccharides and soluble solids were determined in the concentrate for the three extracts. Additionally, the permeate flow, the influences of temperature and pressure, and the resistance to the permeate flow during filtration were also evaluated. A yield of 5.5 g/L of biomass with 35 % glucose conversion was obtained when 0.5 g/L of initial inoculum was employed. Average specific speed of growth was 0.4/day, with biomass productivity of about 0.76 g/(L day). Nanofiltration has yielded polysaccharide increases of 85, 82, and 92 % in the extracts from fruiting bodies, mycelium, and liquid media, respectively. A reduction in the permeate flow was observed during filtration, and it was compensated by higher pressures and temperatures. The higher resistance to the permeate flux was caused by polarization due to concentration (polarized gel layer), reaching values of 88 % for the culture media. Maximal resistance caused by the membrane reached values of 40 % for the extract from the fruiting bodies. On the other hand, resistance caused by fouling was responsible for less than 3.5 %. In conclusion, nanofiltration is efficient to concentrate these functional compounds extracted from A. subrufescens and can, therefore, be applied in different biotechnological areas.     

Seasonal dynamics of Boletus edulis and Lactarius deliciosus extraradical mycelium in pine forests of central Spain

The annual belowground dynamics of extraradical soil mycelium and sporocarp production of two ectomycorrhizal fungi, Boletus edulis and Lactarius deliciosus, have been studied in two different pine forests (Pinar Grande and Pinares Llanos, respectively) in Soria (central Spain). Soil samples (five per plot) were taken monthly (from September 2009 to August 2010 in Pinar Grande and from September 2010 to September 2011 in Pinares Llanos) in eight permanent plots (four for each site). B. edulis and L. deliciosus extraradical soil mycelium was quantified by real-time polymerase chain reaction, with DNA extracted from soil samples, using specific primers and TaqMan® probes. The quantities of B. edulis soil mycelium did not differ significantly between plots, but there was a significant difference over time with a maximum in February (0.1576 mg mycelium/g soil) and a minimum in October (0.0170 mg mycelium/g soil). For L. deliciosus, significant differences were detected between plots and over time. The highest amount of mycelium was found in December (1.84 mg mycelium/g soil) and the minimum in February (0.0332 mg mycelium/g soil). B. edulis mycelium quantities were positively correlated with precipitation of the current month and negatively correlated with the mean temperature of the previous month. Mycelium biomass of L. deliciosus was positively correlated with relative humidity and negatively correlated with mean temperature and radiation. No significant correlation between productivity of the plots with the soil mycelium biomass was observed for any of the two species. No correlations were found between B. edulis sporocarp production and weather parameters. Sporocarp production of L. deliciosus was positively correlated with precipitation and relative humidity and negatively correlated with maximum and minimum temperatures. Both species have similar distribution over time, presenting an annual dynamics characterized by a seasonal variability, with a clear increase on the amounts of biomass during the coldest months of the year. Soil mycelial dynamics of both species are strongly dependent on the weather.     

Morphological and spectrometric analyses of lipids accumulation in a novel oleaginous microalga, Eustigmatos cf. polyphem (Eustigmatophyceae)

Eustigmatos cf. polyphem is a yellow-green unicellular edaphic microalga belonging to the eustigmatophyte. The characteristics of lipids accumulation of E. cf. polyphem grown in a bubble cylindrical photobioreactor under nitrogen-limited conditions was dissected by morphological and spectrometric analyses. Total lipids accumulation rate increased rapidly at early growth phase, with the emergence of many small lipid bodies. Afterwards, lipid bodies became abundant and enlarged primarily because of the progressive accumulation of lipids and the fusion of nearby lipid bodies. Maximum total lipids and neutral lipids content reached up to 60.59 % and 53.08 % of cell dry weight, accompanied with a biomass dry weight 7.9 g/l. E. cf. polyphem is thus referred to as an oleaginous microalga for biodiesel production due to its high biomass and considerable production of oils.     

Medium chain length polyhydroxyalkanoate (mcl-PHA) production from volatile fatty acids derived from the anaerobic digestion of grass

A two step biological process for the conversion of grass biomass to the biodegradable polymer medium chain length polyhydroxyalkanoate (mcl-PHA) was achieved through the use of anaerobic and aerobic microbial processes. Anaerobic digestion (mixed culture) of ensiled grass was achieved with a recirculated leach bed bioreactor resulting in the production of a leachate, containing 15.3 g/l of volatile fatty acids (VFAs) ranging from acetic to valeric acid with butyric acid predominating (12.8 g/l). The VFA mixture was concentrated to 732.5 g/l with a 93.3 % yield of butyric acid (643.9 g/l). Three individual Pseudomonas putida strains, KT2440, CA-3 and GO16 (single pure cultures), differed in their ability to grow and accumulate PHA from VFAs. P. putida CA-3 achieved the highest biomass and PHA on average with individual fatty acids, exhibited the greatest tolerance to higher concentrations of butyric acid (up to 40 mM) compared to the other strains and exhibited a maximum growth rate (μ_MAX?=?0.45 h^?1). Based on these observations P. putida CA-3 was chosen as the test strain with the concentrated VFA mixture derived from the AD leachate. P. putida CA-3 achieved 1.56 g of biomass/l and accumulated 39 % of the cell dry weight as PHA (nitrogen limitation) in shake flasks. The PHA was composed predominantly of 3-hydroxydecanoic acid (>65 mol%).     

Cadmium-induced biochemical responses of Vallisneria spiralis

The following study was carried out to investigate the cadmium (Cd) accumulating potential of Vallisneria. After subjecting plants to different concentrations of Cd, it was observed that plants are able to accumulate ample amount of metal in their roots (5,542 μg g^?1 dw) and leaves (4,368 μg g^?1 dw) in a concentration- and duration-dependent manner. Thus, it is evident that the accumulation in roots was 1.3 times higher than the shoots. It was also noted that with increasing Cd accumulation, roots of the plant appeared darker in color and harder in texture. In response to metal exposure, amount of low molecular weight antioxidants such as cysteine and nonprotein thiols (NP-SH) and activity of enzymes such as APX and GPX were significantly enhanced at lower concentrations of Cd, followed by decline at higher doses. It was also observed that in exposed plants, activity of APX enzyme was higher in roots (ca. 3 times) as compared to leaves. However, chlorophyll and protein content was found to decline significantly in a dose-dependent manner. Results suggested that due to its high accumulation potential, Vallisneria may be effectively grown in water bodies moderately contaminated with Cd.     

Evaluation of Lactobacilli from Human Sources for Uptake and Accumulation of Selenium

The trace mineral selenium (Se) is currently in demand for health supplements for human and animal nutrition. In the present study, 25 isolates of Lactobacillus species of human origin from Indian population were screened for their ability to uptake and accumulate Se in a more bioavailable form. Total Se accumulated by cells was measured by inductively coupled plasma emission spectrometry (ICPES) after wet digestion of Se-enriched cultures. Ten out of 25 isolates grew luxuriantly, as red pigmented colonies, on medium amended with different concentrations of sodium selenite. All the strains when grown in a culture medium supplemented with 10 μg/mL sodium selenite (corresponding to 4.5 μg/mL Se) resulted in Se accumulation in the range of 129.5 to 820.0 μg/g of dry weight (d.w.) as measured using the ICPES method. Elemental Se produced due to reduction of sodium selenite by bacteria was seen as irregular globules under the scanning electron microscope (SEM). Out of all the tested cultures, Lactobacillus reuteri NCDC77 was found to have the greatest ability to uptake Se (28.8 % of inorganic Se in medium) and total Se accumulated was up to 820 μg/g d.w., significantly higher than that of control (742.5 μg/g d.w.). The findings of present study indicate that lactobacilli from human sources have the ability for uptake and accumulation of Se, and the prolific strain has the potential to be explored as an alternative source of organic dietary Se.     

Chemical Composition of the Edible Mushroom Pleurotus ostreatus Produced by Fermentation

Pleurotus ostreatus `Florida' was grown in submerged liquid culture. The biomass yield of the fungus, grown for 3 days in 2-liter fermentors, where the mycelial pellets measuring 5 mm in diameter were formed, was 11.7 g (dry weight)/liter. Comparing the chemical constituents of fruiting bodies produced on cotton straw and mycelial pellets revealed several similarities in total nitrogen, protein, glycogen, fatty acids, RNA, and ash content. Differences were observed in the contents of six amino acids. Although the total fatty acid content was similar, there were more saturated fatty acids in the mycelium. Cell wall composition, typical for basidiomycetes, was observed in both mycelium and fruiting bodies, with laminarin as the main polymer.     

A Study on Cadmium Phytoremediation Potential of Indian Mustard,Brassica juncea

The objective of this study was to investigate Cd phytoremediation ability of Indian mustard, Brassica juncea. The study was conducted with 25, 50, 100, 200 and 400 mg Kg^?1 CdCl₂ in laboratory for 21 days and Cd concentrations in the root, shoot and leaf tissues were estimated by atomic absorption spectroscopy. The plant showed high Cd tolerance of up to 400 mg Kg^?1 but there was a general trend of decline in the root and shoot length, tissue biomass, leaf chlorophyll and carotenoid contents. The tolerance index (TI) of plants were calculated taking both root and shoot lengths as variables. The maximum tolerance (TI _shoot = 87.4 % and TI _root = 89.6 %) to Cd toxicity was observed at 25 mg Kg^?1, which progressively decreased with increase in dose. The highest shoot (10791 μg g^?1 dry wt) and root (9602 μg g^?1 dry wt) Cd accumulation was achieved at 200 mg kg^?1 Cd treatment and the maximum leaf Cd accumulation was 10071.6 μg g^?1 dry wt achieved at 100 mg Kg^?1 Cd, after 21 days of treatment. The enrichment coefficient and root to shoot translocation factor were calculated, which, pointed towards the suitability of Indian mustard for removing Cd from soil.     

Cultivation and utility of <Emphasis Type="Italic">Piptoporus betulinus</Emphasis> fruiting bodies as a source of anticancer agents

Piptoporus betulinus is a wood-rotting basidiomycete used in medicine and biotechnology. However, to date, no indoor method for cultivation of this mushroom fruiting bodies has been developed. Here we present the first report of successful production of P. betulinus mature fruiting bodies in artificial conditions. Four P. betulinus strains were isolated from natural habitats and their mycelia were inoculated into birch sawdust substrate supplemented with organic additives. All the strains effectively colonized the medium but only one of them produced fruiting bodies. Moisture and organic supplementation of the substrate significantly determined the fruiting process. The biological efficiency of the P. betulinus PB01 strain cultivated on optimal substrate (moisture and organic substance content of 55 and 65 and 25 or 35 %, respectively) ranged from 12 to 16 %. The mature fruiting bodies reached weight in the range from 50 to 120 g. Anticancer properties of water and ethanol extracts isolated from both cultured and nature-derived fruiting bodies of P. betulinus were examined in human colon adenocarcinoma, human lung carcinoma and human breast cancer cell lines. The studies revealed antiproliferative and antimigrative properties of all the investigated extracts. Nevertheless the most pronounced effects demonstrated the ethanol extracts, obtained from fruiting bodies of cultured P. betulinus. Summarizing, our studies proved that P. betulinus can be induced to fruit in indoor artificial culture and the cultured fruiting bodies can be used as a source of potential anticancer agents. In this respect, they are at least as valuable as those sourced from nature.     

Cadmium toxicity affects photosynthesis and plant growth at different levels

In this article we discuss and update some of the effects of Cd toxicity on the photosynthetic apparatus in a model crop Lactuca sativa. Seeds of L. sativa were germinated in solutions with 0, 1, 10 and 50 μM of Cd(NO₃)₂ and then transferred to a hydroponic culture medium. After 28 days, the effects of Cd on the photosynthetic apparatus of lettuce were analysed. Exposure of lettuce to 1 μM Cd(NO₃)₂ affected already plant growth (dry biomass), but, did not induce serious damages in the photosynthetic apparatus. However, increasing concentrations of this metal to 10 and 50 μM promoted a strong reduction of the maximum photochemical efficiency of PSII and an impairment of net CO₂ assimilation rate, putatively due to Rubisco activity decrease. This ultimately results in a strong inhibition of plant growth. Nutrient uptake and carbohydrate assimilation were also severely affected by Cd.