Bicarbonate use in three aquatic plants

Our study aimed to test the ability of aquatic plants to use bicarbonate when acclimated to three different bicarbonate concentrations. To this end, we performed experiments with the three species Ceratophyllum demersum, Egeria densa, Lagarosiphon major to determine photosynthetic rates under varying bicarbonate concentrations. We measured bicarbonate use efficiency, photosynthetic performance and respiration. For all species, our results revealed that photosynthetic rates were highest in replicates grown at low alkalinity. Thus, E. densa had approx. five times higher rates at low (264 ± 15 μmol O₂ g⁻¹ DW h⁻¹) than at high alkalinity (50 ± 27 μmol O₂ g⁻¹ DW h⁻¹), C. demersum had three times higher rates (336 ± 95 and 120 ± 31 μmol O₂ g⁻¹ DW h⁻¹), and L. major doubled its rates at low alkalinity (634 ± 114 and 322 ± 119 μmol O₂ g⁻¹ DW h⁻¹). Similar results were obtained for bicarbonate use efficiency by E. densa (136 ± 44 and 43 ± 10 μmol O₂ mequiv. L⁻¹ g⁻¹ DW h⁻¹) and L. major (244 ± 29 and 82 ± 24 μmol O₂ mequiv. L⁻¹ g⁻¹ DW h⁻¹). As to C. demersum, efficiency was high but unaffected by alkalinity, indicating high adaptation ability to varied alkalinities. A pH drift experiment supported these results. Overall, our results suggest that the three globally widespread worldwide species of our study adapt to low inorganic carbon availability by increasing their efficiency of bicarbonate use.     

Characterization of functional intermediates of endoglucanase from Aspergillus aculeatus during urea and guanidine hydrochloride unfolding

Low concentrations of urea and GuHCl (2 M) enhanced the activity of endoglucanase (EC 3.1.2.4) from Aspergillus aculeatus by 2.3- and 1.9-fold, respectively. The K_m values for controls, in the presence of 2 M urea and GuHCl, were found to be 2.4 ± 0.2 × 10⁻⁸ mol L⁻¹, 1.4 ± 0.2 × 10⁻⁸ mol L⁻¹, and 1.6 ± 0.2 × 10⁻⁸ mol L⁻¹, respectively. The dissociation constant (K_d) showed changes in the affinity of the enzyme for the substrate with increases in the K_cat suggesting an increased turnover number in the presence of urea and GuHCl. Fluorescence studies showed changes in the microenvironment of the protein. The increase in the activity of this intermediate state was due to conformational changes accompanied by increased flexibility at the active site.     

Structural, spectroscopic and redox studies of a new ruthenium(III) complex with an imidazole-rich tripodal ligand

The present paper describes a new tripodal ligand containing imidazole and pyridine arms and its first cis-[Ru^III(L)(Cl)₂]ClO₄ complex (1). The crystal structure of 1 shows Ru^III in a distorted octahedral geometry, in which two chloride ions, cis-positioned to each other, are coordinated besides the four nitrogen atoms from the tetradentate ligand L. The cyclic voltammogram of 1 exhibits three redox processes at −67, +73 and +200 mV versus SCE, which are attributed to the Ru^III/Ru^II couple in the cis-[Ru^III(L)(Cl)₂]⁺, cis-[Ru^II(L)(H₂O)(Cl)]⁺ and cis-[Ru^II(L)(H₂O)₂]²⁺, respectively. After chemical reduction (Zn(Hg) or Eu^II) only the cis-[Ru^II(L)(H₂O)₂]²⁺ species is observed in the cyclic voltammetry. Complex 1 absorbs at 470 nm (ε=1.4×10³ mol⁻¹ L cm⁻¹), 335 nm (ε=7.9×10³ mol⁻¹ L cm⁻¹), 301 nm (ε=6.7×10³ mol⁻¹ L cm⁻¹) and 264 nm (ε=9.9×10³ mol⁻¹ L cm⁻¹), in water solution (CF₃COOH, 0.01 mol L⁻¹, μ=0.1 mol L⁻¹ with CF₃COONa). Spectroelectrochemical experiments show a decrease of the bands at 335 and 301 nm, which are attributed to LMCT transitions from the chloride to the Ru^III center and the appearance of a broad band at 402 nm ascribed to MLCT transition from the Ru^II center to the pyridine ligand. The lability of the water ligands in the cis-[Ru^II(L)(H₂O)₂]²⁺ species has been investigated using the auxiliary ligand pyrazine. Reactions in the presence of stoichiometric and excess of pyrazine yield the same species, cis-[Ru^II(L)(H₂O)(pz)]²⁺, which exhibits a reversible redox process at 493 mV versus SCE and absorbs at 438 nm (ε=5.1×10³ mol⁻¹ L cm⁻¹) and 394 nm (ε=4.2×10³ mol⁻¹ L cm⁻¹). Experiments performed with a large excess of pyrazine gave a specific rate constant k₁=(2.8±0.5)×10⁻² M⁻¹ s⁻¹, at 25 °C, in CF₃COOH, 0.01 mol L⁻¹, μ=0.1 mol L⁻¹ (with CF₃COONa).     

Structural and equilibrium studies on mixed ligand complexes of Co(II), Ni(II), Cu(II) and Zn(II) with N-(2-benzimidazolyl)methyliminodiacetic acid and typical N, N donor ligands

Mixed ligand complexes: [Co(L)(bipy)] · 3H₂O (1), [Ni(L)(phen)] · H₂O (2), [Cu(L)(phen)] · 3H₂O (3) and [Zn(L)(bipy)] · 3H₂O (4), where L²⁻ = two -COOH deprotonated dianion of N-(2-benzimidazolyl)methyliminodiacetic acid (H₂bzimida, hereafter, H₂L), bipy = 2,2′ bipyridine and phen = 1,10-phenanthroline have been isolated and characterized by elemental analysis, spectral and magnetic measurements and thermal studies. Single crystal X-ray diffraction studies show octahedral geometry for 1, 2 and 4 and square pyramidal geometry for 3. Equilibrium studies in aqueous solution (ionic strength I = 10⁻¹ mol dm⁻³ (NaNO₃), at 25 ± 1 °C) using different molar proportions of M(II):H₂L:B, where M = Co, Ni, Cu and Zn and B = phen, bipy and en (ethylene diamine), however, provides evidence of formation of mononuclear and binuclear binary and mixed ligand complexes: M(L), M(H₋₁L)⁻, M(B)²⁺, M(L)(B), M(H₋₁L)(B)⁻, M₂(H₋₁L)(OH), (B)M(H₋₁L)M(B)⁺, where H₋₁L³⁻ represents two -COOH and the benzimidazole N₁-H deprotonated quadridentate (O⁻, N, O⁻, N), or, quinquedentate (O⁻, N, O⁻, N, N⁻) function of the coordinated ligand H₂L. Binuclear mixed ligand complex formation equilibria: M(L)(B) + M(B)²⁺ ? (B)M(H₋₁L)M(B)⁺ + H⁺ is favoured with higher π-acidity of the B ligands. For Co(II), Ni(II) and Cu(II), these equilibria are accompanied by blue shift of the electronic absorption maxima of M(II) ions, as a negatively charged bridging benzimidazolate moiety provides stronger ligand field than a neutral one. Solution stability of the mixed ligand complexes are in the expected order: Co(II) < Ni(II) < Cu(II) > Zn(II). The Δ log K_M values are less negetive than their statistical values, indicating favoured formation of the mixed ligand complexes over the binary ones.     

A nanocomposite/crude extract enzyme-based xanthine biosensor

A novel amperometric biosensor for xanthine was developed based on covalent immobilization of crude xanthine oxidase (XOD) extracted from bovine milk onto a hybrid nanocomposite film via glutaraldehyde. Toward the preparation of the film, a stable colloids solution of core–shell Fe₃O₄/polyaniline nanoparticles (PANI/Fe₃O₄ NPs) was dispersed in solution containing chitosan (CHT) and H₂PtCl₆ and electrodeposited over the surface of a carbon paste electrode (CPE) in one step. Scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectrophotometry, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS) were used for characterization of the electrode surface. The developed biosensor (XOD/CHT/Pt NPs/PANI/Fe₃O₄/CPE) was employed for determination of xanthine based on amperometric detection of hydrogen peroxide (H₂O₂) reduction at –0.35 V (vs. Ag/AgCl). The biosensor exhibited a fast response time to xanthine within 8 s and a linear working concentration range from 0.2 to 36.0 μM (R² = 0.997) with a detection limit of 0.1 μM (signal/noise [S/N] = 3). The sensitivity of the biosensor was 13.58 μA μM⁻¹ cm⁻². The apparent Michaelis–Menten (K_m) value for xanthine was found to be 4.7 μM. The fabricated biosensor was successfully applied for measurement of fish and chicken meat freshness, which was in agreement with the standard method at the 95% confidence level.     

Benthic decomposition of Ulva lactuca: A controlled laboratory experiment

The degradation of an Ulva lactuca mat (0.2 kg dw m⁻²) was studied in a controlled flow-through mesocosm for 31 d. Sediment chambers without U. lactuca served as controls. Fluxes of ∑CO₂, O₂, inorganic nitrogen, and urea were determined during the incubation period in addition to sulfate reduction rates, POC and PON content, enumeration of specific bacterial populations and evaluation of the physiological state of the added U. lactuca thalli. After U. lactuca addition to the chambers, there was an immediate increase in the efflux of ∑CO₂ from 11 to 27 mmol-C m⁻² d⁻¹ and a concomitant increase in O₂ uptake from 11 to 23 mmol m⁻² d⁻¹. These effluxes remained elevated throughout the incubation period. In contrast, the NH₄⁺ efflux increased from 0.1 to 1.8 mmol NH₄⁺ m⁻² d⁻¹ during the first 3 d of incubation, followed by 6 d with a constant efflux rate, after which time it decreased gradually to 0.3 mmol NH₄⁺ m⁻² d⁻¹ by the end of the experiment. In total, NH₄⁺accounted for 83% of the total nitrogen efflux after addition of U. lactuca. During the 31 d incubation period there was a continuous colonization of the thalli by bacteria. Sulfate reducers associated with the thalli accounted for 3% of the carbon oxidation on day 31. The molar C:N ratio in mineralization products (the ratio between the efflux of ∑CO₂ and NH₄⁺ + NO₂⁻ + NO₃⁻) increased from 15 mol mol⁻¹ at day 11 after U. lactuca addition to >80 mol mol⁻¹ by the end of the incubation. Since the C:N ratio in the mineralization products was much higher than the original thallus material (8.9 mol mol⁻¹) it is probable that a preferential incorporation of NH₄⁺ into the increasing bacterial biomass occurred. The nitrogen for bacterial growth was most likely obtained from degradation of U. lactuca thalli as there was no stimulation of urea-N turnover in the sediment during incubation. The net increase in bacteria cell number in the 18-mm thick thallus layer was estimated to be 7.6 × 10⁹ to 2.4 × 10¹⁰ bacterial cells cm⁻³. In contrast, the bacterial cell number remained constant in the −Ulva incubations.     

Disposable biosensor and biocatalysis of horseradish peroxidase based on sodium alginate film and room temperature ionic liquid

Composite material based on room temperature ionic liquid (RTIL) N-butylpyridinium hexafluorophosphate (BPPF₆), sodium alginate (SA), and graphite was used to construct a novel horseradish peroxidase (HRP) biosensor for the determination of H₂O₂. The morphology of the as-prepared biosensor was characterized by scanning electron microscopy (SEM). Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to characterize the process of the performance of the biosensor. Parameters affecting the performance of the biosensor, including the concentrations of o-aminophenol (OAP) and HRP and the pH value of substrate solution, were optimized. Under the optimized experimental conditions, H₂O₂ could be detected in a linear calibration range of 1.0 to 6.0 μM with a correlation coefficient of 0.9847 (n = 7) and a detection limit of 0.5 μM at a signal/noise ratio of 3. The prepared biosensor not only had economic and disposable property but also showed good detection precision, bioactivity, storage stability, and reproducibility.     

Chemiluminescent determination of H2O2 using 4-(1,2,4-triazol-1-yl)phenol as an enhancer based on the immobilization of horseradish peroxidase onto magnetic beads

This article describes the employment of a novel p-phenol derivative, 4-(1,2,4-triazol-1-yl)phenol (TRP), as a highly potent signal enhancer of the luminol-hydrogen peroxide (H₂O₂)-horseradish peroxidase (HRP) chemiluminescence (CL) system. The CL reaction conditions were optimized, and the enhancement characteristics of TRP were compared with those of p-iodophenol (PIP). TRP produced a strong enhancement of the CL with the effect of prolonging the light emission. The developed system was then applied to the determination of H₂O₂ with immobilized HRP using magnetic beads as a solid support. The linear range for H₂O₂ was 2.0 × 10⁻⁶ to 1.0 × 10⁻³ M. The detection limit for H₂O₂ was 2.0 × 10⁻⁶ M. The proposed sensor was applied successfully to the determination of H₂O₂ in rainwater.     

Hypoxia and sulphide influence gamete production in Ulva sp.

Gamete production after exposure to hypoxia or sulphide was studied in the marine macroalga Ulva sp. collected in the Sacca di Goro, Italy. Experiments were carried out on discs (12 mm diameter) of thalli cultured in artificial sea water in laboratory at 20 ± 1 °C, 152 μmol m⁻² s⁻¹, 16 h photoperiod and 30‰ salinity. Dehydration of thallus was used as inducer of gametogenesis and growth and gamete release during recovery after 10, 20, 30 or 40 min dehydration (20 ± 1 °C, 25% humidity) were analysed. Unlike non-dehydrated thalli the dehydrated ones produced gametes. Thallus discs, non-dehydrated or subjected to 30 min dehydration, were exposed to hypoxia (1.78–4.02 μmol O₂ L⁻¹) or sulphide (1 mM) for 3, 5, or 7 days at 20 °C in the dark. Non-dehydrated and dehydrated thalli maintained in normoxic conditions in the dark were the controls. Gamete density was checked by counting at the end of the incubation period and during the subsequent 7 days of recovery under 16 h photoperiod in normoxic conditions. Non-dehydrated thalli maintained in normoxic conditions in the dark released gametes when returned to light suggesting that dark constitutes a stimulus to gamete production. The presence of gametes at the end of 3 days incubation of dehydrated thalli in normoxia demonstrated that gametogenesis can occur even in the dark. However, gametes were not present at the end of incubation in hypoxic and sulphidic conditions. Actually, during hypoxic incubation oxygen consumption in D-thalli was very low, only 0.117 × 10⁻³ μmol O₂ mg⁻¹ h⁻¹ compared to 5.93 × 10⁻³ μmol O₂ mg⁻¹ h⁻¹ in normoxia, denoting a reduction of the metabolic rate that could not sustain gametogenesis. During recovery after incubation in normoxic, hypoxic or sulphidic conditions densities of gametes from dehydrated thalli showed significant differences and resulted after hypoxia > after normoxia > after sulphide. Differences in non-dehydrated thalli were not significant. Dehydrated thalli, still green at the end of the incubation period, underwent blanching in the course of recovery in parallel to gamete production, while non-dehydrated thalli maintained their green colour even after exposure to sulphide. Our findings suggest that macroalga Ulva sp. can survive exposure to darkness, severe hypoxia and high sulphide levels and can maintain gamete production even when the exposure to these stress conditions is joined to dehydration.     

Influence of ammonium sulphate feeding time on fed-batch Arthrospira (Spirulina) platensis cultivation and biomass composition with and without pH control

Previous work demonstrated that a mixture of NH₄Cl and KNO₃ as nitrogen source was beneficial to fed-batch Arthrospira (Spirulina) platensis cultivation, in terms of either lower costs or higher cell concentration. On the basis of those results, this study focused on the use of a cheaper nitrogen source mixture, namely (NH₄)₂SO₄ plus NaNO₃, varying the ammonium feeding time (T = 7-15 days), either controlling the pH by CO₂ addition or not. A. platensis was cultivated in mini-tanks at 30 °C, 156 μmol photons m⁻² s⁻¹, and starting cell concentration of 400 mg L⁻¹, on a modified Schlösser medium. T = 13 days under pH control were selected as optimum conditions, ensuring the best results in terms of biomass production (maximum cell concentration of 2911 mg L⁻¹, cell productivity of 179 mg L⁻¹ d⁻¹ and specific growth rate of 0.77 d⁻¹) and satisfactory protein and lipid contents (around 30% each).     

Light-dependent phosphate uptake of a submersed macrophyte Myriophyllum spicatum L.

The uptake kinetics of phosphate (Pi) by Myriophyllum spicatum was determined from adsorption and absorption under light and dark conditions. Pi uptake was light dependent and showed saturation following the Michaelis-Menten relation (in light: V = 16.91 × [Pi](1.335 + [Pi]), R² = 0.90, p < 0.001; in the dark: V = 5.13 × [Pi](0.351 + [Pi]), R² = 0.77, p < 0.001). Around 77% of the loss of Pi in the water column was absorbed into the tissue of M. spicatum, and only 23% was adsorbed on the surface of the plant shoots. Our study shows that M. spicatum shoots have a much higher affinity (in light: 3.9 μmol g⁻¹ dw h⁻¹ μM⁻¹; in the dark: 3.7 μmol g⁻¹ dw h⁻¹ μM⁻¹) and V_max (maximum uptake rate, shoot light) for Pi uptake than many other aquatic macrophytes (in light: 0.002-0.23 μmol g⁻¹ dw h⁻¹ μM⁻¹; in the dark: 0.002-0.19 μmol g⁻¹ dw h⁻¹ μM⁻¹), which may provide a competitive advantage over other macrophytes across a wide range of Pi concentrations.     

Development of cesium phosphotungstate salt and chitosan composite membrane for direct methanol fuel cells

A novel composite membrane has been developed by doping cesium phosphotungstate salt (Cs_xH_3−xPW₁₂O₄₀ (0 ≤ x ≤3), Cs_x-PTA) into chitosan (CTS/Cs_x-PTA) for application in direct methanol fuel cells (DMFCs). Uniform distribution of Cs_x-PTA nanoparticles has been achieved in the chitosan matrix. The proton conductivity of the composite membrane is significantly affected by the Cs_x-PTA content in the composite membrane as well as the Cs substitution in PTA. The highest proton conductivity for the CTS/Cs_x-PTA membranes was obtained with x = 2 and Cs₂-PTA content of 5 wt%. The value is 6 × 10⁻³ S cm⁻¹ and 1.75 × 10⁻² S cm⁻¹ at 298 K and 353 K, respectively. The methanol permeability of CTS/Cs₂-PTA membrane is about 5.6 × 10⁻⁷, 90% lower than that of Nafion-212 membrane. The highest selectivity factor (φ) was obtained on CTS/Cs₂-PTA-5 wt% composite membrane, 1.1 × 10⁴/S cm⁻³ s. The present study indicates the promising potential of CTS/Cs_x-PTA composite membrane as alternative proton exchange membranes in direct methanol fuel cells.     

Water column anammox and denitrification in a temperate permanently stratified lake (Lake Rassnitzer,Germany)

We studied microbial N₂ production via anammox and denitrification in the anoxic water column of a restored mining pit lake in Germany over an annual cycle. We obtained high-resolution hydrochemical profiles using a continuous pumping sampler. Lake Rassnitzer is permanently stratified at ca. 29 m depth, entraining anoxic water below a saline density gradient. Mixed-layer nitrate concentrations averaged ca. 200 μmol L⁻¹, but decreased to zero in the anoxic bottom waters. In contrast, ammonium was <5 μmol L⁻¹ in the mixed layer but increased in the anoxic waters to ca. 600 μmol L⁻¹ near the sediments. In January and October, ¹⁵N tracer measurements detected anammox activity (maximum 504 nmol N₂ L⁻¹ d⁻¹ in ¹⁵NH₄⁺-amended incubations), but no denitrification. In contrast, in May, N₂ production was dominated by denitrification (maximum 74 nmol N₂ L⁻¹ d⁻¹). Anammox activity in May was significantly lower than in October, as characterized by anammox rates (maximum 6 vs. 16 nmol N₂ L⁻¹ d⁻¹ in incubations with ¹⁵NO₃⁻), as well as relative and absolute anammox bacterial cell abundances (0.56% vs. 0.98% of all bacteria, and 2.7×10⁴ vs. 5.2×10⁴ anammox cells mL⁻¹, respectively) (quantified by catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH) with anammox bacteria-specific probes). Anammox bacterial diversity was investigated with anammox bacteria-specific 16S rRNA gene clone libraries. The majority of anammox bacterial sequences were related to the widespread Candidatus Scalindua sorokinii/brodae cluster. However, we also found sequences related to Candidatus S. wagneri and Candidatus Brocadia fulgida, which suggests a high anammox bacterial diversity in this lake comparable with estuarine sediments.     

Hydrolytic cleavage of double-strand DNA by the water-soluble dicobalt(III) complexes of 1,4,7-Triazacyclononane-N-acetate

Three water-soluble dicobalt(III) complexes, [Co₂L₂(µ-OH)₂](ClO₄)₂·5H₂O (1), [Co₂L₂(µ-OH)₂](ClO₄)₂·CH₃OH·H₂O(2); [Co₂L₂(µ-OH)₂](ClO₄)₂·4H₂O(3) (L = 1,4,7-triazacyclononane-N-acetate monoanion), were prepared to serve as nuclease mimics. The complexes were characterized by X-ray, IR and UV-vis spectroscopy as well as ESI-MS. Three complexes exhibit similar structures, just with different solvent molecules. The electrospray mass spectrum of 1 in solution indicates that dinuclear ion [Co₂L₂(µ-OH)₂-H⁺] ⁺ (4) is the active species. In the absence of any reducing agent, the complexes cleave plasmid pBR322 DNA was performed and its hydrolytic mechanism was demonstrated with radical scavengers, anaerobic reaction and T4 ligase. The kinetic aspects of DNA cleavage under pseudo- or true-Michaelis-Menten conditions are also detailed, kinetic parameters (k_cat, K_M) were calculated to be 3.57 h^− 1, 6.92 × 10^− 4 M; 0.28 h^− 1, 1.9 × 10^− 5 M for 4, respectively.     

UV-Vis and fluorimetric Al, Zn, Cd and Pb complexation studies of two 3-hydroxyflavones and a 3-hydroxythioflavone

The complexation of Al³⁺, Zn²⁺, Cd²⁺ and Pb²⁺ by the 3-hydroxyflavones: 3-hydroxy-2-(2-methoxyphenyl)-4H-1-benzopyran-4-one (H1) and 3-hydroxy-2-(4-methoxyphenyl)-4H-1-benzopyran-4-one (H2), and by the 3-methoxythioflavone: 3-hydroxy-2-(2-methoxyphenyl)-4H-1-benzopyran-4-thione (H3) have been studied spectrophotometrically and fluorimetrically to determine the corresponding complexation constants, K_sp and K_fl, in 5:95 water:ethanol (v/v) solution for which [HClO₄] was either 10⁻² or 10⁻⁵ mol dm⁻³ and I = 0.10 mol dm⁻³ (NaClO₄) at 298.2 K. Complexation occurs dominantly through the deprotonated ligand for [Al(1)]²⁺ and [Al(2)]²⁺ for which log K_sp = 4.51 and 4.73, respectively, in 10⁻² mol dm⁻³ HClO₄ and 4.21 and 4.61 in 10⁻⁵ mol dm⁻³ HClO₄. For Pb²⁺ complexation by H1, H2 and H3 is characterized by log K_sp = 2.20, 2.57 and 3.22, respectively, in 10⁻² mol dm⁻³ HClO₄ and 4.70, 5.38 and 5.74 in 10⁻⁵ mol dm⁻³ HClO₄. Equilibrium mixtures of [Pb(H1)]²⁺ and [Pb1]⁺, [Pb(H2)]²⁺ and [Pb2]⁺, and [Pb(H3)]²⁺ and [Pb3]⁺ appear to be formed. Complexation of Zn²⁺ and Cd²⁺ by all three ligands was only detected in 10⁻⁵ mol dm⁻³ HClO₄. For Zn²⁺ complexation by H1, H2 and H3 log K_sp = 3.22, 3.74 and 4.46 and for Cd²⁺ the corresponding values are 2.39, 2.40 and 3.72 for Cd²⁺. Only [Al1]²⁺ and [Al2]²⁺ show significant fluorescence and are characterized by log K_fl = 6.30 and 7.49 in 10⁻² mol dm⁻³ HClO₄.     

Nitrogen nutrition of Canna indica: Effects of ammonium versus nitrate on growth, biomass allocation, photosynthesis, nitrate reductase activity and N uptake rates

The effects of inorganic nitrogen (N) source (NH₄⁺, NO₃⁻ or both) on growth, biomass allocation, photosynthesis, N uptake rate, nitrate reductase activity and mineral composition of Canna indica were studied in hydroponic culture. The relative growth rates (0.05-0.06 g g⁻¹ d⁻¹), biomass allocation and plant morphology of C. indica were indifferent to N nutrition. However, NH₄⁺ fed plants had higher concentrations of N in the tissues, lower concentrations of mineral cations and higher contents of chlorophylls in the leaves compared to NO₃⁻ fed plants suggesting a slight advantage of NH₄⁺ nutrition. The NO₃⁻ fed plants had lower light-saturated rates of photosynthesis (22.5 μmol m⁻² s⁻¹) than NH₄⁺ and NH₄⁺/NO₃⁻ fed plants (24.4-25.6 μmol m⁻² s⁻¹) when expressed per unit leaf area, but similar rates when expressed on a chlorophyll basis. Maximum uptake rates (V_max) of NO₃⁻ did not differ between treatments (24-35 μmol N g⁻¹ root DW h⁻¹), but V_max for NH₄⁺ was highest in NH₄⁺ fed plants (81 μmol N g⁻¹ root DW h⁻¹), intermediate in the NH₄NO₃ fed plants (52 μmol N g⁻¹ root DW h⁻¹), and lowest in the NO₃⁻ fed plants (28 μmol N g⁻¹ root DW h⁻¹). Nitrate reductase activity (NRA) was highest in leaves and was induced by NO₃⁻ in the culture solutions corresponding to the pattern seen in fast growing terrestrial species. Plants fed with only NO₃⁻ had high NRA (22 and 8 μmol NO₂⁻ g⁻¹ DW h⁻¹ in leaves and roots, respectively) whereas NRA in NH₄⁺ fed plants was close to zero. Plants supplied with both forms of N had intermediate NRA suggesting that C. indica takes up and assimilate NO₃⁻ in the presence of NH₄⁺. Our results show that C. indica is relatively indifferent to inorganic N source, which together with its high growth rate contributes to explain the occurrence of this species in flooded wetland soils as well as on terrestrial soils. Furthermore, it is concluded that C. indica is suitable for use in different types of constructed wetlands.     

Large catalase based bioelectrode for biosensor application

A large catalase (CAT) (Mr ~ 90 kDa), immobilized on multiwalled carbon nanotubes—Nafion^® (MWCNT-NF) matrix and encapsulated with polyethylenimine (PEI) on glassy carbon electrode (GCE), showed a pair of nearly reversible cyclic voltammetric peaks for Fe^(III)/Fe^(II) couple with formal potential of about −0.45 V (vs. Ag/AgCl electrode at pH 7.5). PEI significantly reduced the charge transfer resistance and stabilized the bioelectrode through electrostatic interaction. The electron transfer rate constant and surface coverage of the immobilized CAT were 1.05 ± 0.2 s⁻¹ and 2.1 × 10⁻¹⁰ mol cm⁻², respectively. Studies on electrocatalytic activity and kinetics of GCE/MWCNT-NF/CAT/PEI for hydrogen peroxide (H₂O₂) showed the apparent Michaelis-Menten constant of 3 mM, linear response in the range of 10 μM to 5 mM, response time of ~ 2 s for steady state current, and detection limit of ~ 1 μM. A high operational and storage stability was also demonstrated for the bioelectrode. Hence, the direct electrochemistry of the large catalase and its potential biosensor application have been established through this investigation.     

Morphological and physiological adaptive traits of Mediterranean narrow endemic plants: The case of Centaurea gymnocarpa (Capraia Island,Italy)

A case study on Centaurea gymnocarpa Moris & De Not., a narrow endemic species, was carried out by analyzing its morphological, anatomical, and physiological traits in response to natural habitat stress factors under Mediterranean climate conditions. The results underline that the species is particularly adapted to the environment where it naturally grows. At the plant level, the above-ground/below-ground dry mass (1.73 ± 0.60) shows its investment predominately in the above-ground structure with a resulting total leaf area per plant of 1399 ± 94 cm². The senescent attached leaves at the base of the plant contribute to limit leaf transpiration by shading soil around the plant. Moreover, the dense C. gymnocarpa leaf pubescence, leaf rolling, the relatively high leaf mass area (LMA = 12.3 ± 1.3 mg cm⁻²) and leaf tissue density (LTD = 427 ± 44 mg cm⁻³) contribute to limit leaf transpiration, also postponing leaf death under dry conditions. At the physiological level, a relatively low respiration/photosynthesis ratio (R/P_N) in spring results from high R [2.26 ± 0.59 μmol (CO₂) m⁻² s⁻¹] and P_N [12.3 ± 1.5 μmol (CO₂) m⁻² s⁻¹]. The high photosynthetic nitrogen use efficiency [PNUE = 15.5 ± 0.4 μmol (CO₂) g⁻¹ (N) s⁻¹] shows the large amount of nitrogen (N) invested in the photosynthetic machinery of new leaves, associated to a high chlorophyll content (Chl = 35 ± 5 SPAD units). On the contrary, the highest R/P_N ratio (1.75 ± 0.19) in summer is due to a significant P_N decrease and increase of R in response to drought. The low PNUE [1.5 ± 0.2 μmol (CO₂) g⁻¹ (N) s⁻¹] in this season is indicative of a greater N investment in leaf cell walls which may contribute to limit transpiration. On the contrary, the low R/P_N ratio (0.05 ± 0.02) in winter is resulting from the limited enzyme activity of the respiratory apparatus [R = 0.23 ± 0.08 μmol (CO₂) m⁻² s⁻¹] while the low PNUE [3.5 ± 0.2 μmol (CO₂) g⁻¹ (N) s⁻¹] suggests that low temperatures additionally limit plant production. The experiment of the imposed water stress confirms that the C. gymnocarpa growth capability is in conformity with the severe conditions of its natural habitat, likewise as it may be the case with others narrow endemic species that have occupied niches with similar extreme conditions.     

Short term effects of hypoxia and bioturbation on solute fluxes, denitrification and buffering capacity in a shallow dystrophic pond

The effects of short term hypoxia on bioturbation activity and inherent solute fluxes are scarcely investigated even if increasing number of coastal areas are subjected to transient oxygen deficits. In this work dark fluxes of oxygen (O₂), dissolved inorganic carbon (TCO₂) and nutrients across the sediment-water interface, as well as rates of denitrification (isotope pairing), were measured in intact sediment cores collected from the dystrophic pond of Sali e Pauli (Sardinia, Italy). Sediments were incubated at 100, 70, 40 and 10% of O₂ saturation in the overlying water, with both natural benthic communities, dominated by the polychaete Polydora ciliata (11.100 ± 2.500  ind. m^− 2), and after the addition of individuals of the deep-burrower polychaete Hediste diversicolor. Below an uppermost oxic layer of ~ 1 mm, sediments were highly reduced, with up to 6 mM of S²⁻ in the 5 mm layer. Flux of S²⁻ and O₂ calculated from pore water gradients were 8.61 ± 1.12 and − 2.27 ± 0.56 mmol m^− 2 h^− 1, respectively. However, sediment oxygen demand (SOD) calculated from core incubation was − 10.52 ± 0.33 mmol m^− 2 h^− 1, suggesting a major contribution of P. ciliata to O₂-mediated sulphide oxidation. P. ciliata also strongly stimulated NH₄⁺ and PO₄³⁻ fluxes, with rates ~ 15 and ~ 30 folds higher, respectively, than those estimated from pore water gradients. P. ciliata activity was significantly reduced at 10% O₂ saturation, coupled to decreased rates of solutes transfer. The addition of H. diversicolor further stimulated SOD, NH₄⁺ efflux and SiO₂ mobilisation. Similarly to P. ciliata, the degree of stimulation of SOD and NH₄⁺ flux by H. diversicolor depended on the level of oxygen saturation. TCO₂ regeneration, respiratory quotients, PO₄³⁻ fluxes and denitrification of added ¹⁵NO₃⁻ were not affected by the addition of H. diversicolor, but depended upon the O₂ levels in the water column. Denitrification rates supported by water column ¹⁴NO₃⁻ and sedimentary nitrification were both negligible (< 0.5 µmol m^− 2 h^− 1). They were not significantly affected by oxygen saturation nor by bioturbation, probably due to the limited availability of NO₃⁻ in the water column (< 3 µM) and O₂ in the sediments. This study demonstrates for the first time the integrated short term effect of transient hypoxia and bioturbation on solute fluxes across the sediment-water interface within a simplified lagoonal benthic community.