Production, Optimization, Characterization and Drought Stress Resistance by β-Glucan-Rich Heteropolysaccharide From an Endophytic Fungi Colletotrichum alatae LCS1 Isolated From Clubmoss (Lycopodium clavatum)

Endophytic entities are ubiquitous in nature with all-square bioactivity ranging from therapeutic effects toward animals to growth promoting attributes and stress tolerance activities in case of green plants. In the present study, the club moss Lycopodium clavatum for the first time has been subjected for the isolation of endophytic fungi. An exopolysaccharide (EPS) extracted from Colletotrichum alatae LCS1, an endophytic fungi isolated from L. clavatum Linn., was characterized as a β-glucan heteropolymer (composed of mannose, rhamnose, arabinose, glucose, galactose, and fucose) which plays a pivotal role in obliterating the drought stress in rice seedlings (Oryza sativa) when applied at an amount of 20, 50, and 100 ppm. The fresh weight contents of rice tissue (39%), total chlorophyll (33%), proline (41%), soluble sugar content (26%) along with antioxidant enzymes such as catalase, peroxidase, and super-oxide dismutase increased (in comparison to control of non-EPS treated seedlings) while malondialdehyde content had reduced markedly after 30 days of regular treatment. The drought resistance of rice seedling was observed at peak when applied at 50 ppm dosage. Vital parameters for EPS production like fermentation duration (5 days), medium pH (6), nutrient (carbon (glucose-7 g%/l), nitrogen (yeast extract-0.4 g%/l), and mineral (NaCl-0.10 g%/l) sources, oxygen requirements (O2 vector or liquid alkane-n-hexane, n-heptane, n-hexadecane), and headspace volume (250 ml Erlenmeyer flask- 50 ml medium, 200 ml-headspace volume) were optimized to obtain an enhanced EPS yield of 17.38 g/L−59% higher than the preoptimized one. The present study, for the first time, reported the β-glucan rich heteropolysaccharide from Colletotrichum origin which is unique in structure and potent in its function of drought stress tolerance and could enhance the sustainable yield of rice cultivation in areas facing severe drought stress.

Impedance Spectroscopy applied to the study of high dilutions of Lycopodium clavatum

Introduction: The Impedance spectroscopy [1] is a technique mainly used to characterize the electrical behavior of solids or liquids samples. This particular technique involves placing the sample of material under investigation between two electrodes (capacitor plates), applying an AC voltage and observing the resulting response across the spectrum of impedance by plotting the real part (Z’) as a function of the imaginary part (Z”) of the impedance. Alternatively, graphs of either the real or the imaginary parts of the impedance can be constructed as a function of the applied voltage frequency. Comparative measurements previously carried out by Miranda et al [2]. have demonstrated clear differences between the impedance values of high dilutions of lithium chloride (LiCl) and the corresponding reference water samples (water which has undergone the same dinamization procedures but without the salt). In this paper the results obtained by applying the spectroscopy of impedance technique in high dilutions of Lycopodium clavatum - Lyc (from 15cH to 30 cH), in comparison to the reference waters, will be presented and discussed. Aims: The objective of this work is to measure the impedance components of both high dilutions of Lycopodium clavatum and reference water samples in the frequency range of 100Hz to 13Mhz, using a successful protocol of sample preparation which has already been used before2. Details of the experimental set-up can be found elsewhere[3]. Methodology: Thirty samples of Lyc solutions and thirty reference water samples were produced using the same preparation and measuring protocol. Both groups of liquid samples were measured for dynamizations ranging from 1cH to 30cH, in accordance to the Hahnemanian dynamization method and following the practice suggested by the Brazilian Homeopathic Pharmacopeia. The Lyc solutions were specifically compared to the reference water samples in the potencies of 15cH, 18cH, 23cH and 30cH. It is important to highlight here that all the Lyc solutions and the corresponding reference water samples measured were prepared from the same lot of initial distilled water and submitted to the same steps of dilution and succussion protocol3. Typically three impedance measurements were carried out for each investigated solution, starting with the highest potency. The sample holder (capacitor cell) used during the experiment was careful and systematically cleaned after each measurement. Results: The results obtained show that by choosing either the real part (Z’) or the imaginary component (Z”) of the impedance, it is possible to clearly differentiate the Lyc solutions from the corresponding reference water samples, for the potencies 15cH, 18cH and 30cH. For the potency 23cH, however, this difference is not very significant, as it can be observed in Figure 1. Conclusion: Impedance spectroscopy has demonstrated itself to be a powerful and sensitive technique for the physical characterization of Lycopodium clavatum in high dilutions. The differences obtained for the LiCl dynamizations and the corresponding pure water samples are noteworthy. Also, the results exhibit a non-monotonic behavior over the process of dynamization, indicating that the possibility of contamination during the samples manipulation can be ruled out.

Causticum hahnemanni, Conium maculatum and Lycopodium clavatum highly diluted medications decreases parasitemia in mice infected by Trypanosoma cruzi

Introduction: Benznidazole is the only medicine available in Brazil for Chagas’ disease treatment, however it presents low efficacy in the chronic phase and several adverse effects [1-3]. Aim: Evaluate the effect of Causticum hahnemanni, Conium maculatum and Lycopodium clavatum [4-6] administered to mice infected with T. cruzi. Method: In blind randomized controlled trial 42 male Swiss mice, 8 weeks of age, have been grouped: GCaus –treated with C. hahnemanni 13cH (n=10), GCon –treated with C. maculatum 13 cH (n=11), GLy –treated with L. clavatum13cH (n=10) and CG – control group (n=11) treated with 7% hydro alcoholic solution 13cH. The animals were infected intraperitoneally with 1.400 blood trypomastigotes of T. cruzi - Y strain. Medications were been prepared according to Brazilian Homeopathic Pharmacopoeia [7]. Medication was diluted in water (1mL/100mL) offered ad libitum, from amber recipient during 16 hours administered 48 hours before infection and 48, 96 and 144 hours after inoculation. Parasitological parameters assessed: total parasitemia (TP), maximum peak of parasites (MPP), pre-patent period (PPP) and area under curve (AUC). Parasitemia was evaluated daily counting from the first day of infection. Clinical parameters assessed: weight, temperature, water and food intake and excreta were measured counting from the 5º day before infection until animals’ death or checking negative parasitemia for 3 consecutive days. Mortality was registered for 75 days after infection. Ethics Committee for Experiments in Animals gave approval UEM 054/11. Statistical comparison of data was performed with Kruskal-Wallis test, with 5% significance. Results and Discussion: The diluted medications have been significantly reduced the parasitological parameters: MPP (p

Soil Pollen Indicators of Historical Arable Horizons from the Zvenigorod Biological Station of the Moscow State University

In this work we present the results of spore and pollen analysis of forest soils from the Zvenigorod biological station of Moscow State University (Moscow Region, Russia). A comparative analysis of forest soils formed on the site of historical fields of the XVIII–XIX centuries and beyond showed that a specific complex of pollen and spores remains in the residual arable horizons, characteristic only of soils that have passed through the stages of plowing and fallow. It includes pollen from cultivated cereals and arable weeds (buckwheat, cornflower blue), spores of the mace-shaped plaunus (Lycopodium clavatum), as well as spores of the mosses Riccia glauca and Anthoceros spp. The latter are exclusive indicators of fallows, since they are practically not found in other habitats. The identified pollen indicators can be used in landscape and archaeological research to interpret the data of spore-pollen analysis of cultural layers, buried soils, gully-ravine sediments. They can also be used to define the boundaries of ancient fields under modern vegetation.

Influence of the Pyrotechnic Igniter Composition Aging on Explosion Parameters of Dispersed Dusts

A commercially available pyrotechnic igniter was used according to the EN 14034 and ASTM E1226a Standards to study the explosiveness of dispersed dusts. Its pyrotechnic composition consists of 1.2 g of zirconium (40% wt.), barium peroxide (30% wt.) and barium nitrate (30% wt.). The energy released during the combustion of that amount of composition is 5 kJ. The article investigates the influence of aging of the pyrotechnic composition in the igniter on its initiation parameters. In the study, igniters of different years from date of manufacture were used: Igniter 1, manufactured in 2021 (less than 1 year from date of manufacture), and Igniter 2 (more than 2 years from date of manufacture). The study was performed in the KV 150M2 explosion chamber with a volume of 365 L and the 20 L sphere chamber with a volume of 20 L. A standard sample of Lycopodium clavatum was used in the KV 150M2 explosion chamber. Magnesium and benzoic acid were used as the samples in the 20 L sphere explosion chamber. The experiment showed that the explosion pressure Pmax of the igniter with more than 2 years from date of manufacture decreased by up to 10%, while the value of the explosion constant Kst decreased by up to 40%. The attained results proved that aging of igniters affects their explosion parameters and measurement accuracy.

The Toxicity Evaluation of Lycopodiacea Extracts by Mice Neural Cell and Zebrafish Embryo

Many species in the family Lycopodiacea possess highly potential medical substances, notably are Huperzine A of Huperzia serrata and alkaloids isolated from Lycopodium clavatum(α-Onocerin , Lycopodine,..). Various members of this family are found throughout of Vietnam and a few researches on its diversity as well as alkaloid isolation has been conducted, however, there is a lack of toxicology studies on this subject. In this study, three crude extracts (labeled AI.1, AI.2 and AI.3), which come from Lycopodium sp.collected from 3 different regions of Vietnam, have been tested on 2 toxicological models: primary neural cells derived from Swiss white mouse (in vitro) and Zebrafish (Danio rerio) embryos (in vivo). Our results show that the all three extracts are relatively low in toxicity, IC50 value on primary neural cell ranging from 800 - 1100 mg/L, while LC50 of fish embryos at 100 - 300 mg/L. AI.2 has the least effect of viability of both cells and embryos compare to the others. This data give the prove showed that the safety using Lycopodium as tradional medicine.

The Uptake of Sporopollenin Exine Capsules and Associated Bioavailability of Adsorbed Oestradiol in Selected Aquatic Invertebrates

Lycopodium clavatum sporopollenin exine capsules (SpECs) are known to both adsorb and absorb chemicals. The aim of the present work was to determine whether oestradiol (E2) is ‘bioavailable’ to bioindicator species, either pre-adsorbed to, or in the presence of, SpECs. SpEC uptake was confirmed for Daphnia magna and Dreissena bugensis. E2 levels varied among treatments for Caenorhabditis elegans though there was no relationship to SpEC load. E2 was not detected in D. bugensis tissues. Expression changes of general stress and E2-specific genes were measured. For C. elegans, NHR-14 expression suggested that SpECs modulate E2 impacts, but not general health responses. For D. magna, SpECs alone and with E2 changed Vtg1 and general stress responses. For D. bugensis, SpECS were taken up but no E2 or change in gene expression was detected after exposure to E2 and/or SpECs. The present study is the first to investigate SpECs and bound chemical dynamics.