Nitrogen Depletion and Redistribution by Free‐Ranging Cattle in the Restoration Process of Mosaic Landscapes: The Role of Foraging Strategy and Habitat Proportion

In a mosaic landscape in N‐Belgium (W‐Europe), consisting of forest, grassland, and wooded pasture on former agricultural land, we assessed nitrogen redistribution by free‐ranging cattle (±0.2 animal units ha^?1 yr^?1). We examined if the spatial redistribution of nitrogen among habitats by cattle could restore nutrient‐poor conditions in preferred foraging habitats, and conversely whether such translocation could lead to extreme eutrophication in preferred resting habitats. We used nitrogen content of different diet classes, habitat use, foraging and defecation behavior, weight gain, and nitrogen losses in the actual situation to explore four different habitat proportion scenarios and two different foraging strategies to calculate a net nitrogen balance per habitat. An atmospheric deposition of 30 kg N ha^?1 yr^?1 with varying interception factors according to the habitat types was included in an integrated nitrogen balance. All scenarios showed a net nitrogen transport from grassland and wooded pasture to forest habitat. We found that nitrogen redistribution strongly depends on habitat proportion. Nitrogen losses from preferred grassland habitat can be high, given its proportion is small. Depletion is only to be expected at excretion‐free areas and probably is of minor importance to trigger the establishment of woody species. In general, nitrogen transported by cattle was much lower than input by atmospheric deposition, but grazing can compensate for high N inputs in excretion‐free areas and maintain grassland types that support critical loads of 20–25 kg N ha^?1 yr^?1. In none of the scenarios, N transport by cattle resulted in the exceeding of critical nitrogen loads to vulnerable forest ground vegetation.     

Biodegradation of Shrimp Biowaste by Marine Exiguobacterium sp. CFR26M and Concomitant Production of Extracellular Protease and Antioxidant Materials: Production and Process Optimization by Response Surface Methodology

Twelve marine bacterial cultures were screened for extracellular protease activity, and the bacterium CFR26M which exhibited the highest activity on caseinate agar plate was identified as an Exiguobacterium sp. Significant amount of extracellular protease (5.9?±?0.3 U/ml) and antioxidant materials, measured as 2,2′-diphenyl picrylhydrazyl (DPPH) radical scavenging activity (44.4?±?0.5 %), was produced by CFR26M in submerged fermentation using a shrimp biowaste medium. Response surface methodology (RSM) was employed to optimize the process variables for maximum production of protease and antioxidant materials by CFR26M. Among the seven variables screened by two-level 2**(7–2) fractional factorial design, the concentration of shrimp biowaste, sugar, and phosphate was found to be significant (p?≤?0.05). The optimum levels of these variables were determined by employing the central composite design (CCD) of RSM. The coefficient of determination (R ²) values of 0.9039 and 0.8924 for protease and antioxidant, respectively, indicates the accuracy of the CCD models. The optimum levels of shrimp biowaste, sugar, and phosphate were 21.2, 10.5, and 2.3 % (w/v) for production of protease and 28.8, 12, and 0.32 % (w/v) for production of antioxidant material, respectively. The concentration of shrimp biowaste, sugar, and phosphate had linear and quadratic effect on both protease and antioxidant productions. RSM optimization yielded 6.3-fold increases in protease activity and 1.6-fold in antioxidant material production. The crude protease of CFR26M had a maximum activity at 32?±?2 °C with pH 7.6. This is the first report on the use of marine Exiguobacterium sp. for concomitant production of protease and antioxidant materials from shrimp biowaste.     

Remediation of Gulf War Oil Spill Contaminated Soil by a Subcritical Water Extraction Process: Oil Removal,Recovery, and Degradation

Due to the unique properties of subcritical water (marked change in water's dielectric constant and viscosity), the extraction by subcritical water offers a great opportunity to remediate soil contaminated with organic pollutants as an alternative and green remediation method. In this study, subcritical water extraction is proposed as an efficient remediation technique for the Gulf War oil spill contaminated soil. The subcritical water extraction experiment was carried out in a lab-scale continuous flow apparatus. The three major operating factors, temperature, time and water flow rate, were evaluated in terms of optimum removal efficiency. The results show that crude oil removal depended largely on water temperature, whereas an extraction run time higher than 1 h and a water flow rate higher than 1.5 mL/min marginally or negatively affected removal efficiency. During subcritical water treatment at 300°C for 1 h at a flow rate of 1.5 mL/min, removal efficiency was almost 95%. Under these operating conditions, the subcritical water treatment demonstrated a similar removal efficiency to those of organic solvents like acetone. In contrast, the efficiency of oil recovery decreased with an increase in extraction temperature, due to degradation by a water self-oxidizing agent. Several degradation products identified in the treated soil and in the effluent sample (which initially were absent in the contaminated soil) were oxygen-containing aromatic compounds, confirming the oxidation-degradation.     

Attraction of Mexican Fruit Flies (Diptera: Tephritidae) to Grapefruit: Enhancement by Mechanical Wounding of and Experience with Grapefruit

Wild strain, mated, female Mexican fruit flies, Anastrepha ludens (Loew), with no prior experience with fruit (naive), were not attracted to grapefruit, a preferred cultivated host, in wind tunnel experiments. Naive, mated laboratory strain females were attracted. Prior experience with grapefruit increased attraction of both laboratory and wild strains. More females were attracted to fruit with peel damage than to undamaged fruit, indicating that fruit odor mediated attraction. More naive than experienced females attempted to oviposit on the sides of the wind tunnel. Experienced laboratory males, but not wild males or naive males, were attracted to grapefruit. Attraction and oviposition behavior of laboratory flies was greater than that of wild flies.     

Genetic Variation of Morphological Traits and Transpiration in an Apple Core Collection under Well-Watered Conditions: Towards the Identification of Morphotypes with High Water Use Efficiency

Water use efficiency (WUE) is a quantitative measurement which improvement is a major issue in the context of global warming and restrictions in water availability for agriculture. In this study, we aimed at studying the variation and genetic control of WUE and the respective role of its components (plant biomass and transpiration) in a perennial fruit crop. We explored an INRA apple core collection grown in a phenotyping platform to screen one-year-old scions for their accumulated biomass, transpiration and WUE under optimal growing conditions. Plant biomass was decompose into morphological components related to either growth or organ expansion. For each trait, nine mixed models were evaluated to account for the genetic effect and spatial heterogeneity inside the platform. The Best Linear Unbiased Predictors of genetic values were estimated after model selection. Mean broad-sense heritabilities were calculated from variance estimates. Heritability values indicated that biomass (0.76) and WUE (0.73) were under genetic control. This genetic control was lower in plant transpiration with an heritability of 0.54. Across the collection, biomass accounted for 70% of the WUE variability. A Hierarchical Ascendant Classification of the core collection indicated the existence of six groups of genotypes with contrasting morphology and WUE. Differences between morphotypes were interpreted as resulting from differences in the main processes responsible for plant growth: cell division leading to the generation of new organs and cell elongation leading to organ dimension. Although further studies will be necessary on mature trees with more complex architecture and multiple sinks such as fruits, this study is a first step for improving apple plant material for the use of water.     

Complexes of (dG-dC)20 with Mn2+ Ions: A Study by Vibrational Circular Dichroism and Infrared Absorption Spectroscopy

Abstract

The B-Z transition of the synthetic oligonucleotide, (dG-dC)₂₀, induced by Mn²⁺ ions at room temperature, was investigated by absorption and Vibrational Circular Dichroism (VCD) spectroscopy in the range of 1800–800 cm^?1. Metal ion concentration was varied from 0 to 0.73 M Mn²⁺ (0 to 8.5 moles of Mn²⁺ per mole of oligonucleotide phosphate, [Mn]/[P]). While both types of spectra showed considerable changes as the Mn²⁺ concentrations were raised, differences between the two were often complementary in their expression and extent, those displayed by VCD being more clearly evident due to the inversion of the opposite helical sense from the right-handed to the left-handed conformation. The main phase of the transition occurred in the metal ion concentration between 0.8?1.1 [Mn]/[P]. Gradual changes that took place in the spectra were interpreted in terms of simultaneous processes that depended on metal ion concentration, namely B-Z transformation, binding of Mn²⁺ to phosphates and to nitrogen bases, and partial denaturation. Below～0.6 [Mn]/[P], only a small portion of the oligonucleotide adopted the Z conformation within a 3 hour period, whereas conversion was completed in the same time interval for concentrations between 0.9?1.2 [Mn]/[P]. At [Mn]/[P] > 1.7, complete transition to the Z-form took place immediately on adding Mn²⁺. Applying VCD spectroscopy in combination with conventional infrared absorption proved most useful for corroborating changes in the absorption spectra, and for detecting in an unique manner, not attainable by absorption methods, conformational changes that lead to the inversion of the helical sense of the oligonucleotide.     

Efficiency of mitochondrial uncoupling by modified butyltriphenylphosphonium cations and fatty acids correlates with lipophilicity of cations: Protonophoric vs leakage mechanisms

In order to determine the share of protonophoric activity in the uncoupling action of lipophilic cations a number of analogues of butyltriphenylphosphonium with substitutions in phenyl rings (C₄TPP-X) were studied on isolated rat liver mitochondria and model lipid membranes. An increase in the rate of respiration and a decrease in the membrane potential of isolated mitochondria were observed for all the studied cations, the efficiency of these processes was significantly enhanced in the presence of fatty acids and correlated with the octanol-water partition coefficient of the cations. The ability of C₄TPP-X cations to induce proton transport across the lipid membrane of liposomes loaded with a pH-sensitive fluorescent dye increased also with their lipophilicity and depended on the presence of palmitic acid in the liposome membrane. Of all the cations, only butyl[tri(3,5-dimethylphenyl)]phosphonium (C₄TPP-diMe) was able to induce proton transport by the mechanism of formation of a cation-fatty acid ion pair on planar bilayer lipid membranes and liposomes. The rate of oxygen consumption by mitochondria in the presence of C₄TPP-diMe increased to the maximum values corresponding to conventional uncouplers; for all other cations the maximum uncoupling rates were significantly lower. We assume that the studied cations of the C₄TPP-X series, with the exception of C₄TPP-diMe at low concentrations, cause nonspecific leak of ions through lipid model and biological membranes which is significantly enhanced in the presence of fatty acids.     

Rapid Healing of Cutaneous Leishmaniasis by High-Frequency Electrocauterization and Hydrogel Wound Care with or without DAC N-055: A Randomized Controlled Phase IIa Trial in Kabul

Background

Anthroponotic cutaneous leishmaniasis (CL) due to Leishmania (L.) tropica infection is a chronic, frequently disfiguring skin disease with limited therapeutic options. In endemic countries healing of ulcerative lesions is often delayed by bacterial and/or fungal infections. Here, we studied a novel therapeutic concept to prevent superinfections, accelerate wound closure, and improve the cosmetic outcome of ACL.

Methodology/Principal Findings

From 2004 to 2008 we performed a two-armed, randomized, double-blinded, phase IIa trial in Kabul, Afghanistan, with patients suffering from L. tropica CL. The skin lesions were treated with bipolar high-frequency electrocauterization (EC) followed by daily moist-wound-treatment (MWT) with polyacrylate hydrogel with (group I) or without (group II) pharmaceutical sodium chlorite (DAC N-055). Patients below age 5, with facial lesions, pregnancy, or serious comorbidities were excluded. The primary, photodocumented outcome was the time needed for complete lesion epithelialization. Biopsies for parasitological and (immuno)histopathological analyses were taken prior to EC (1^st), after wound closure (2^nd) and after 6 months (3^rd). The mean duration for complete wound closure was short and indifferent in group I (59 patients, 43.1 d) and II (54 patients, 42 d; p = 0.83). In patients with Leishmania-positive 2^nd biopsies DAC N-055 caused a more rapid wound epithelialization (37.2 d vs. 58.3 d; p = 0.08). Superinfections occurred in both groups at the same rate (8.8%). Except for one patient, reulcerations (10.2% in group I, 18.5% in group II; p = 0.158) were confined to cases with persistent high parasite loads after healing. In vitro, DAC N-055 showed a leishmanicidal effect on pro- and amastigotes.

Conclusions/Significance

Compared to previous results with intralesional antimony injections, the EC plus MWT protocol led to more rapid wound closure. The tentatively lower rate of relapses and the acceleration of wound closure in a subgroup of patients with parasite persistence warrant future studies on the activity of DAC N-055.

Trial Registration

ClinicalTrails.gov {"type":"clinical-trial","attrs":{"text":"NCT00947362","term_id":"NCT00947362"}}NCT00947362     

Remediation of Urban River Water by Pontederia Cordata Combined with Artificial Aeration: Organic Matter and Nutrients Removal and Root-Adhered Bacterial Communities

Macrophyte combined with artificial aeration is a promising in situ remediation approach for urban rivers polluted with nutrients and organic matter. However, seasonal variations and aeration effects on phytoremediation performance and root-adhered microbial communities are still unclear. In this study, Pontederia cordata was used to treat polluted urban river water under various aeration intensities. Results showed that the highest removal efficiencies of chemical oxygen demand (COD_Cr) and total nitrogen (TN) were attained under aeration of 30 L min^?1 in spring and summer and 15 L min^?1 in autumn, while total phosphorus (TP) removal reached maximum with aeration of 15 L min^?1 in all seasons. Moderate aeration was beneficial for increasing the diversity of root-adhered bacteria communities, and the shift of bacterial community structure was more pronounced in spring and autumn with varying aeration intensity. The dual effect, i.e. turbulence and dissolved oxygen (DO), of aeration on the removal of COD_Cr and TN prevailed over the individual effect of DO, while DO was the most influential factor for TP removal and the root-adhered bacterial community diversity. P. cordata combined with 15 L min^?1 aeration was deemed to be the best condition tested in this study.     

Biosynthesis of Abscisic Acid in the Fungus Cercospora cruenta: Stimulation of Biosynthesis by Water Stress and Isolation of a Transgenic Mutant with Reduced Biosynthetic Capacity

The rate of synthesis of abscisic acid (ABA) by the phytopathogenicfungus Cercospora cruenta was found to be increased 2.6-foldby water stress (0.5 M mannitol, 48 h). To characterize theoxidative steps in the synthesis of ABA by C. cruenta, a mutantstrain deficient in the synthesis of ABA was isolated aftertransformation with a foreign gene. This strain was furthercharacterized by an immunoassay using a monoclonal antibodyspecific for 1',4'-dihydroxy--ionylidene-acetate. One transformantthat seemed likly to be deficient in the synthesis of ABA wasisolated. Integration of the foreign gene into the chromosomalDNA of the transformant was confirmed by Southern hybridization. ³Present address: Nitto Chemical Industries Co. Ltd.Chiyodaku,Tokyo, 100 Japan     

On the Permeation by Dioxygen of Urate Oxidase from Aspergillus flavus in Complex with Xanthine Anion: Dioxygen Pathways and a Portrait of the Enzyme Cavities from Molecular Dynamics Simulations in Water Solution

This work describes molecular dynamics (MD) simulations in aqueous media for the complex of the homotetrameric urate oxidase (UOX) from Aspergillus flavus with xanthine anion ( 5 ) in the presence of dioxygen (O₂). After 196.6 ns of trajectory from unrestrained MD, a O₂ molecule was observed leaving the bulk solvent to penetrate the enzyme between two subunits, A/C. From here, the same O₂ molecule was observed migrating, across subunit C, to the hydrophobic cavity that shares residue V227 with the active site. The latter was finally attained, after 378.3 ns of trajectory, with O₂ at a bonding distance from 5 . The reverse same O₂ pathway, from 5 to the bulk solvent, was observed as preferred pathway under random acceleration MD (RAMD), where an external, randomly oriented force was acting on O₂. Both MD and RAMD simulations revealed several cavities populated by O₂ during its migration from the bulk solvent to the active site or backwards. Paying attention to the last hydrophobic cavity that apparently serves as O₂ reservoir for the active site, it was noticed that its volume undergoes ample fluctuations during the MD simulation, as expected from the thermal motion of a flexible protein, independently from the particular subunit and no matter whether the cavity is filled or not by O₂.