Effects of wood char and activated carbon on the hydrolysis of cellobiose by β-glucosidase from Aspergillus niger

Biochar amendments to soils have been suggested as a strategy to sequester carbon and therefore mitigate global climate change. The enrichment of soils with charred materials also increases their fertility. This fertilising effect of biochar may be caused by various mechanisms; an acceleration of nutrient cycling has been suggested as one such mechanism. The rate-limiting step in nutrient cycling is thought to be the extracellular enzymatic attack on biological macromolecules. In this study, therefore, the effects of chestnut wood char (specific surface area 2.0 m² g⁻¹) and of activated carbon (specific surface area approximately 900 m² g⁻¹) on an extracellular enzymatic reaction involved in the degradation of cellulose (i.e., hydrolysis of cellobiose by β-glucosidase from Aspergillus niger) were investigated. Cellobiose was not adsorbed by chestnut wood char, whereas activated carbon absorbed more than 97% of it. Both charred materials adsorbed more than 99% of β-glucosidase. For chestnut wood char, adsorption of the enzyme caused a decrease of approximately 30% in the reaction rate, whereas for activated carbon, the nearly complete absorption of both substrate and enzyme entirely inhibited the reaction. These results show that β-glucosidase from A. niger retains most of its activity when adsorbed to chestnut wood char and that the reaction it catalyses in nature is only slightly affected by this charred material. On the other hand, a material characterised by a high specific surface area and high porosity, such as activated carbon, can make even a highly soluble substrate unavailable for soil enzymes and therefore completely inhibit the reaction. Thus, charred materials may affect nutrient cycling mainly by regulating the availability of substrates: the degradation of highly soluble substrates may be accelerated by materials with low specific surface area, which maintain an active and protected enzyme pool, whereas materials with high specific surface and high porosity may slow down the degradation by making substrates unavailable.     

Brain proton magnetic resonance spectroscopy findings in a Beagle dog with genetically confirmed Lafora disease

Cortical atrophy has been identified using magnetic resonance imaging (MRI) in humans and dogs with Lafora disease (LD). In humans, proton magnetic resonance spectroscopy (1HMRS) of the brain indicates decreased N-acetyl-aspartate (NAA) relative to other brain metabolites. Brain 1HMRS findings in dogs with LD are lacking. A 6-year-old female Beagle was presented with a history of a single generalized tonic-clonic seizure and episodic reflex myoclonus. Clinical, hematological, and neurological examination findings and 3-Tesla MRI of the brain were unremarkable. Brain 1HMRS with voxel positioning in the thalamus was performed in the affected Beagle. It identified decreased amounts of NAA, glutamate-glutamine complex, and increased total choline and phosphoethanolamine relative to water and total creatine compared with the reference range in healthy control Beagles. A subsequent genetic test confirmed LD. Abnormalities in 1HMRS despite lack of changes with conventional MRI were identified in a dog with LD.     

Earthworms can modify effects of hydrochar on growth of Plantago lanceolata and performance of arbuscular mycorrhizal fungi

Hydrothermal carbonization (HTC) is a method to produce carbonized material at relatively low temperatures (180–250 °C) under pressure and aqueous conditions. The product is called hydrochar and can be used as a soil amendment. However, applied in high dosages it may have detrimental effects on plants or soil biota. The potential impact of hydrochar amendment on beneficial soil organisms such as arbuscular mycorrhizal fungi (AMF) and earthworms and their interactions are not well understood. The goal of the present study was to determine effects of hydrochar on plant growth and soil biota and to evaluate interactions of earthworms and hydrochar on plant and AMF performance and to identify underlying mechanisms. In a greenhouse experiment, we investigated the effect of hydrochar at different addition rates (control, 1% and 10%, v/v) with or without the earthworm Aporrectodea caliginosa on the growth of Plantago lanceolata L. and the performance of its AMF. We observed a positive interaction between earthworms and 10% hydrochar on shoot and root biomass: added as a single treatment hydrochar had a negative effect on plant growth at this dosage, but plant biomass increased significantly when hydrochar was added together with earthworms. Root colonization by AMF increased significantly with increasing concentration of hydrochar, but was not affected by earthworms. Contrastingly, extraradical hyphal length of AMF was reduced by earthworms, but not affected by hydrochar. Thus, hydrochar and earthworms affected the performance of AMF, albeit of different AMF structures and in different directions. Our results indicate that earthworms may play an important role in alleviating the negative impacts of high dosages of hydrochar on plant growth; such interactions should move into focus of future research on potential effects of HTC materials.     

Isolation and characterization of a Mn(II)-oxidizing Bacillus strain from the demosponge Suberites domuncula

In this study we demonstrate that the demosponge Suberites domuncula harbors a Mn(II)-oxidizing bacterium, a Bacillus strain, termed BAC-SubDo-03. Our studies showed that Mn(II) stimulates bacterial growth and induces sporulation. Moreover, we show that these bacteria immobilize manganese on their cell surface. Comparison of the 16S rDNA sequence allowed the grouping of BAC-SubDo-03 to the Mn-precipitating bacteria. Analysis of the spore cell wall revealed that it contains an Mn(II)-oxidizing enzyme. Co-incubation studies of BAC-SubDo-03 with 100 μM MnCl(2) and >1 μM of CuCl(2) showed an increase in their Mn(II)-oxidizing capacity. In order to prove that a multicopper oxidase-like enzyme(s) (MCO) exists in the cell wall of the S. domuncula-associated BAC-SubDo-03 Bacillus strain, the gene encoding this enzyme was cloned (mnxG-SubDo-03). Sequence alignment of the deduced MCO protein (MnxG-SubDo-03) revealed that the sponge bacterium clusters together with known Mn(II)-oxidizing bacteria. The expression of the mnxG-SubDo-03 gene is under strong control of extracellular Mn(II). Based on these findings, we assume that BAC-SubDo-03 might serve as a Mn reserve in the sponge providing the animal with the capacity to detoxify Mn in the environment. Applying the in vitro primmorph cell culture system we could demonstrate that sponge cells, that were co-incubated with BAC-SubDo-03 in the presence of Mn(II), show an increased proliferation potential.     

Echinacea alkylamides modulate induced immune responses in T-cells

The effects of Echinacea and several of its phytochemical components on NFkappaB expression by Jurkat cells (a human T-cell line) were investigated in vitro. In the absence of stimulation, Echinacea and its components exerted no significant effect on basal NFkappaB expression levels. In the presence of endotoxin (LPS), NFkappaB expression was decreased. However, this decrease was significantly reversed by treatment with cichoric acid, an Echinacea root extract (prepared from both Echinacea angustifolia and Echinacea purpurea) and the alkylamide fraction derived from this combination. For the phorbol myristate acetate stimulation of Jurkat cells, effects on NFkappaB expression were mixed. Depending on the concentration, cichoric acid and a 2,4-diene alkylamide significantly induced NFkappaB levels, whereas a 2-ene alkylamide caused a significant inhibition. In contrast, both the Echinacea and the mixed alkylamide fraction exerted no effect. The alkylamide results indicate that the two basic forms of these compounds present in Echinacea may have opposing effects. These opposing effects demonstrate the importance of a knowledge, not only of the phytochemical make-up of a herbal preparation, but also of the actions of each component and the consequences of differing relative amounts in the preparation being investigated.     

Flux partitioning in an old-growth forest: seasonal and interannual dynamics

Turbulent fluxes of carbon, water and energy were measured at the Wind River Canopy Crane, Washington, USA from 1999 to 2004 with eddy-covariance instrumentation above (67 m) and below (2.5 m) the forest canopy. Here we present the decomposition of net ecosystem exchange of carbon (NEE) into gross primary productivity (GPP), ecosystem respiration (R(eco)) and tree canopy net CO(2) exchange (DeltaC) for an old-growth Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco)-western hemlock (Tsuga heterophylla (Raf.) Sarg.) forest. Significant amounts of carbon were recycled within the canopy because carbon flux measured at the below-canopy level was always upward. Maximum fluxes reached 4-6 micromol m(-2) s(-1) of CO(2) into the canopy air space during the summer months, often equaling the net downward fluxes measured at the above-canopy level. Ecosystem respiration rates deviated from the expected exponential relationship with temperature during the summer months. An empirical ecosystem stress term was derived from soil water content and understory flux data and was added to the R(eco) model to account for attenuated respiration during the summer drought. This attenuation term was not needed in 1999, a wet La Ni?a year. Years in which climate approximated the historical mean, were within the normal range in both NEE and R(eco), but enhanced or suppressed R(eco) had a significant influence on the carbon balance of the entire stand. In years with low respiration the forest acts as a strong carbon sink (-217 g C m(-2) year(-1)), whereas years in which respiration is high can turn the ecosystem into a weak to moderate carbon source (+100 g C m(-2) year(-1)).     

Seasonal change in the drought response of wood cell development in poplar

Field-grown poplar trees (Populus nigra L. x P. maximowiczii Henry, clone Kamabuchi) were exposed to severe drought twice during the growing season to evaluate the impact on wood cell development. The drought treatment caused a reduction in leaf water potential, leaf wilting and a decreased concentration of osmotically active solutes in the cambial zone. Drought-induced changes in the anatomy of developing xylem cells were examined in stem sections and macerated wood samples. In early summer, drought significantly reduced the length and cross-sectional area of newly formed fibers, whereas no such effects were observed in late summer. In well-watered trees, fiber cross-sectional area declined between early and late summer. Similarly, drought reduced the cross-sectional area of vessel elements in early summer but not in late summer, whereas in both control and drought-treated trees, the cross-sectional area of vessel elements decreased between early and late summer. The vessel area to xylem area ratio was unaffected by drought because the drought-induced decrease in vessel size was matched by an increase in the number of newly formed vessel cells. In contrast to its effect in early summer, late-summer drought had no significant effect on fiber and vessel cell development, indicating that sensitivity of wood cell development to drought varies seasonally.     

Segregation of microsatellite loci in second generation hybrid striped bass (<Emphasis Type="Italic">Morone saxatilis</Emphasis> × <Emphasis Type="Italic">Morone chrysops</Emphasis>)

Within the framework of a larger project aimed to assess the potential of second generation hybrid striped bass for German aquaculture the genotypic segregation of five microsatellite loci was analysed in two progeny lots (n = 74 and 76, respectively). There was no consistent correlation between microsatellite genotypes and phenotypic category (white bass, hybrid, or striped bass). None of the individuals expressed neither only white bass nor only striped bass genotypes at all five loci. On the other hand, only hybrid genotypes at all five loci were detected in three individuals of lot 1 and four individuals of lot 2. Single loci tests for conformity of microsatellite genotypic segregation with Mendelian rules revealed significant deviations (P < 0.05) in four cases for lot 1 and in three cases for lot 2. If pooled over all five loci, both lots displayed highly significant deviations (P < 0.01) with an excess of hybrid genotypes and a deficiency of white bass genotypes. It is concluded that stabilizing selection performed on hybrid genotypes might be a suitable approach for practical aquaculture in Europe if the goal is to become independent of first generation hybrid fry supply and/or if establishing domesticated brood stocks of both parental species is impossible. However, more detailed studies on the characteristics and performance of multiple hybrid generations are needed.     

An unexpected host for the endangered giant freshwater pearl mussel Margaritifera auricularia (Spengler, 1793) as a conservation tool

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