Nitrogen uptake kinetics in field populations and cultured strains of Karenia brevis

This study represents the most comprehensive assessment of kinetic parameters for Karenia brevis to date as it encompasses natural populations sampled during three different bloom years in addition to cultured strains under controlled conditions. Nitrogen (N) uptake kinetics for ammonium (NH₄⁺), nitrate (NO₃⁻), urea, an amino acid mixture, individual amino acids (glutamate and alanine), and humic substrates were examined for the toxic red tide dinoflagellate, K. brevis, during short term incubations (0.5–1 h) using ¹⁵N tracer techniques. Experiments were conducted using natural populations collected during extensive blooms along the West Florida Shelf in October 2001, 2002, and 2007, and in cultured strains (CCFWC 251 and CCFWC 267) obtained from the Florida Fish and Wildlife Institute culture collection. Kinetic parameters for the maximum uptake velocity (V_max), half-saturation concentration (K_s), and the affinity constant (α) were determined. The affinity constant is considered a more accurate indicator of substrate affinity at low concentrations. K. brevis took up all organic substrates tested, including N derived from humic substances. Uptake rates of the amino acid mixture and some NO₃⁻ incubations did not saturate even at the highest substrate additions (50–200 μmol N L⁻¹). Based upon the calculated α values, the greatest substrate preference was for NH₄⁺ followed by NO₃⁻ ≥ urea, humic compounds and amino acids. The ability of K. brevis to utilize a variety of inorganic and organic substrates likely helps it flourish under a wide range of nutrient conditions from bloom initiation in oligotrophic waters offshore to bloom maintenance near shore where ambient nutrient concentrations may be orders of magnitude greater.     

Detection of Karenia brevis blooms on the west Florida shelf using in situ backscattering and fluorescence data

Using shipboard data collected from the central west Florida shelf (WFS) between 2000 and 2001, an optical classification algorithm was developed to differentiate toxic Karenia brevis blooms (>10⁴ cells l⁻¹) from other waters (including non-blooms and blooms of other phytoplankton species). The identification of K. brevis blooms is based on two criteria: (1) chlorophyll a concentration ≥1.5 mg m⁻³ and (2) chlorophyll-specific particulate backscattering at 550 nm ≤ 0.0045 m² mg⁻¹. The classification criteria yielded an overall accuracy of 99% in identifying both K. brevis blooms and other waters from 194 cruise stations. The algorithm was validated using an independent dataset collected from both the central and south WFS between 2005 and 2006. After excluding data from estuarine and post-hurricane turbid waters, an overall accuracy of 94% was achieved with 86% of all K. brevis bloom data points identified successfully. Satisfactory algorithm performance (88% overall accuracy) was also achieved when using underway chlorophyll fluorescence and backscattering data collected during a repeated alongshore transect between Tampa Bay and Florida Bay in 2005 and 2006. These results suggest that it may be possible to use presently available, commercial optical backscattering instrumentation on autonomous platforms (e.g. moorings, gliders, and AUVs) for rapid and timely detection and monitoring of K. brevis blooms on the WFS.     

Nitrogen fixation and methanotrophy in forest mosses along a N deposition gradient

Nitrogen deposition has decreased the plant-associated nitrogen (N₂) fixation when measured using the indirect acetylene reduction assay (ARA). However, nitrogen deposition can also lead to changes in the diversity of moss symbionts, e.g. affect methanotrophic N₂ fixation, which is not measured by ARA. To test this hypothesis we compared ARA with the direct stable isotope method (¹⁵N₂ incorporation) and studied methanotrophy in two mosses, Hylocomium splendens and Pleurozium schreberi, collected from seven forest sites along a boreal latitudinal N deposition transect. We recognized that the two independent N₂ fixation measures gave corresponding results with the conversion factor of 3.3, but the ¹⁵N₂ method was more sensitive for finding a signal of low N₂ fixation activity. Methane carbon fixation associated with mosses was under the detection limit (<2 nmol C g⁻¹ h⁻¹). N₂ fixation rates were more pronounced in the mosses with higher C/N ratio, and in the green upper parts of the shoot than in the lower brownish parts. Sequencing of nifH genes revealed that dominating diazotrophs were affiliated to cyanobacterial genera Nostoc and Nodularia, but methanotrophic diazotrophs were not found in the nifH libraries. We conclude that the suppression of N₂ fixation along the deposition gradient was consistent regardless of the measurement technique, and microbial community changes toward methanotrophic or otherwise acetylene-sensitive N₂ fixation could not explain this trend.     

Influence of daylight surface aggregation behavior on nutrient cycling during a Karenia brevis (Davis) G. Hansen & Ø. Moestrup bloom: Migration to the surface as a nutrient acquisition strategy

The toxic HAB dinoflagellate Karenia brevis (Davis) G. Hansen & Ø. Moestrup (formerly Gymnodinium breve) exhibits a migratory pattern atypical of dinoflagellates: cells concentrate in a narrow (∼0–5 cm) band at the water surface during daylight hours due to phototactic and negative geotactic responses, then disperse downward at night via non-tactic, random swimming. The hypothesis that this daylight surface aggregation behavior significantly influences bacterial and algal productivity and nutrient cycling within blooms was tested during a large, high biomass (chlorophyll a >19 μg L⁻¹) K. brevis bloom in October of 2001 by examining the effects of this surface layer aggregation on inorganic and organic nutrient concentrations, cellular nitrogen uptake, primary and bacterial productivity and the stable isotopic signature (δ¹⁵N, δ¹³C) of particulate material. During daylight hours, concentrations of K. brevis and chlorophyll a in the 0–5 cm surface layer were enhanced by 131% (±241%) and 32.1% (±86.1%) respectively compared with an integrated water sample collection over a 0–1 m depth. Inorganic (NH₄, NO₃₊₂, PO₄, SiO₄) and organic (DOP, DON) nutrient concentrations were also elevated within the surface layer as was both bacterial and primary productivity. Uptake of nitrogen (NH₄⁺, NO₃⁻, urea, dissolved primary amines, glutamine and alanine) compounds by K. brevis was greatest in the surface layer for all compounds tested, with the greatest enhancement evident in urea uptake rates, from 0.08 × 10⁻⁵ ng N K. brevis cell⁻¹ h⁻¹ to 3.1 × 10⁻⁵ ng N K. brevis cell⁻¹ h⁻¹. These data suggests that this surface aggregation layer is not only an area of concentrated cells within K. brevis blooms, but also an area of increased biological activity and nutrient cycling, especially of nitrogen. Additionally, the classic dinoflagellate migration paradigm of a downward migration for access to elevated NO₃⁻ concentrations during the dark period may not apply to certain dinoflagellates such as K. brevis in oligotrophic nearshore areas with no significant nitricline. For these dinoflagellates, concentration within a narrow surface layer in blooms during daylight hours may enhance nutrient supply through biological cycling and photochemical nutrient regeneration.     

Discrimination against 15N among recombinant inbred lines of Phaseolus vulgaris L. contrasting in phosphorus use efficiency for nitrogen fixation

Although isotopic discrimination processes during nitrogen (N) transformations influence the outcome of ¹⁵N based quantification of N₂ fixation in legumes, little attention has been given to the effects of genotypic variability and environmental constraints such as phosphorus (P) deficiency, on discrimination against ¹⁵N during N₂ fixation. In this study, six Phaseolus vulgaris recombinant inbred lines (RILs), i.e. RILs 115, 104, 34 (P deficiency tolerant) and 147, 83, 70 (P deficiency sensitive), were inoculated with Rhizobium tropici CIAT899, and hydroaeroponically grown with P-sufficient (250 μmol P plant⁻¹ week⁻¹) versus P-deficient (75 μmol P plant⁻¹ week⁻¹) supply. Two harvests were done at 15 (before nodule functioning) and 42 (flowering stage) days after transplanting. Nodulation, plant biomass, P and N contents, and the ratios of ¹⁵N over total N content (¹⁵N/Nt) for shoots, roots and nodules were determined. The results showed lower ¹⁵N/Nt in shoots than in roots, both being much lower than in nodules. P deficiency caused a larger decrease in ¹⁵N/Nt in shoots (−0.18%) than in nodules (−0.11%) for all of the genotypes, and the decrease in shoots was greatest for RILs 34 (−0.33%) and 104 (−0.25%). Nodule ¹⁵N/Nt was significantly related to both the quantity of N₂ fixed (R² = 0.96***) and the P content of nodules (R² = 0.66*). We conclude that the discrimination against ¹⁵N in the legume N₂-fixing symbiosis of common bean with R. tropici CIAT899 is affected by P nutrition and plant genotype, and that the ¹⁵N/Nt in nodules may be used to screen for genotypic variation in P use efficiency for N₂ fixation.     

Pea plant responsiveness under elevated [CO2] is conditioned by the N source (N2 fixation versus NO3− fertilization)

The main goal of this study was to test the effect of [CO₂] on C and N management in different plant organs (shoots, roots and nodules) and its implication in the responsiveness of exclusively N₂-fixing and NO₃⁻-fed plants. For this purpose, exclusively N₂-fixing and NO₃⁻-fed (10 mM) pea (Pisum sativum L.) plants were exposed to elevated [CO₂] (1000 μmol mol⁻¹ versus 360 μmol mol⁻¹ CO₂). Gas exchange analyses, together with carbohydrate, nitrogen, total soluble proteins and amino acids were determined in leaves, roots and nodules. The data obtained revealed that although exposure to elevated [CO₂] increased total dry mass (DM) in both N treatments, photosynthetic activity was down-regulated in NO₃⁻-fed plants, whereas N₂-fixing plants were capable of maintaining enhanced photosynthetic rates under elevated [CO₂]. In the case of N₂-fixing plants, the enhanced C sink strength of nodules enabled the avoidance of harmful leaf carbohydrate build up. On the other hand, in NO₃⁻-fed plants, elevated [CO₂] caused a large increase in sucrose and starch. The increase in root DM did not contribute to stimulation of C sinks in these plants. Although N₂ fixation matched plant N requirements with the consequent increase in photosynthetic rates, in NO₃⁻-fed plants, exposure to elevated [CO₂] negatively affected N assimilation with the consequent photosynthetic down-regulation.     

Nitrogen uptake and regeneration (ammonium regeneration,nitrification and photoproduction) in waters of the West Florida Shelf prone to blooms of Karenia brevis

The West Florida Shelf (WFS) encompasses a range of environments from inshore estuarine to offshore oligotrophic waters, which are frequently the site of large and persistent blooms of the toxic dinoflagellate, Karenia brevis. The goals of this study were to characterize the nitrogen (N) nutrition of plankton across the range of environmental conditions on the WFS, to quantify the percentage of the plankton N demand met through in situ N regeneration, and to determine whether planktonic N nutrition changes when high concentrations of Karenia are present. In the fall of 2007, 2008, and 2009 we measured ambient nutrient concentrations and used stable isotope techniques to measure rates of primary production and uptake rates of inorganic N (ammonium, NH₄⁺, and nitrate, NO₃⁻), and organic N and carbon (C; urea and amino acids, AA) in estuarine, coastal, and offshore waters, as well as coastal sites with Karenia blooms present. In parallel, we also measured rates of in situ N regeneration – NH₄⁺ regeneration, nitrification, and photoproduction of NH₄⁺, nitrite and AA. Based on microscope observations, ancillary measurements, and previous monitoring history, Karenia blooms sampled represented three bloom stages – initiation in 2008, maintenance in 2007, and late maintenance/stationary phase in 2009. Nutrient concentrations were highest at estuarine sampling sites and lowest at offshore sites. Uptake of NH₄⁺ and NO₃⁻ provided the largest contribution to N nutrition at all sites. At the non-Karenia sites, in situ rates of NH₄⁺ regeneration and nitrification were generally sufficient to supply these substrates equal to the rates at which they were taken up. At Karenia sites, NO₃⁻ was the most important N substrate during the initiation phase, while NH₄⁺ was the most important N form used during bloom maintenance and stationary phases. Rates of NH₄⁺ regeneration were high but insufficient (85 ± 36% of uptake) to support the measured NH₄⁺ uptake at all the Karenia sites although nitrification rates far exceeded uptake rates of NO₃⁻. Taken together our results support the “no smoking gun” nutrient hypothesis that there is no single nutrient source or strategy that can explain Karenia's frequent dominance in the waters where it occurs. Consistent with other papers in this volume, our results indicate that Karenia can utilize an array of inorganic and organic N forms from a number of N sources.     

Soil nitrate accumulation explains the nonlinear responses of soil CO2 and CH4 fluxes to nitrogen addition in a temperate needle-broadleaved mixed forest

The responses of soil-atmosphere carbon (C) exchange fluxes to growing atmospheric nitrogen (N) deposition are controversial, leading to large uncertainty in the estimated C sink of global forest ecosystems experiencing substantial N inputs. However, it is challenging to quantify critical load of N input for the alteration of the soil C fluxes, and what factors controlled the changes in soil CO₂ and CH₄ fluxes under N enrichment. Nine levels of urea addition experiment (0, 10, 20, 40, 60, 80, 100, 120, 140 kg N ha⁻¹ yr⁻¹) were conducted in the needle-broadleaved mixed forest in Changbai Mountain, Northeast China. Soil CO₂ and CH₄ fluxes were monitored weekly using the static chamber and gas chromatograph technique. Environmental variables (soil temperature and moisture in the 0–10 cm depth) and dissolved N (NH₄⁺-N, NO₃⁻-N, total dissolved N (TDN), and dissolved organic N (DON)) in the organic layer and the 0–10 cm mineral soil layer were simultaneously measured. High rates of N addition (≥60 kg N ha⁻¹ yr⁻¹) significantly increased soil NO₃⁻-N contents in the organic layer and the mineral layer by 120%-180% and 56.4%-84.6%, respectively. However, N application did not lead to a significant accumulation of soil NH₄⁺-N contents in the two soil layers except for a few treatments. N addition at a low rate of 10 kg N ha⁻¹ yr⁻¹ significantly stimulated, whereas high rate of N addition (140 kg N ha⁻¹ yr⁻¹) significantly inhibited soil CO₂ emission and CH₄ uptake. Significant negative relationships were observed between changes in soil CO₂ emission and CH₄ uptake and changes in soil NO₃⁻-N and moisture contents under N enrichment. These results suggest that soil nitrification and NO₃⁻-N accumulation could be important regulators of soil CO₂ emission and CH₄ uptake in the temperate needle-broadleaved mixed forest. The nonlinear responses to exogenous N inputs and the critical level of N in terms of soil C fluxes should be considered in the ecological process models and ecosystem management.     

Proton production from nitrogen transformation drives stream export of base cations in acid-sensitive forested watersheds

The biogeochemical cycles of nitrogen (N) and base cations (BCs), (i.e., K⁺, Na⁺, Ca²⁺, and Mg²⁺), play critical roles in plant nutrition and ecosystem function. Empirical correlations between large experimental N fertilizer additions to forest ecosystems and increased BCs loss in stream water are well demonstrated, but the mechanisms driving this coupling remain poorly understood. We hypothesized that protons generated through N transformation (PPR_N)—quantified as the balance of NH₄⁺ (H⁺ source) and NO₃⁻ (H⁺ sink) in precipitation versus the stream output will impact BCs loss in acid-sensitive ecosystems. To test this hypothesis, we monitored precipitation input and stream export of inorganic N and BCs for three years in an acid-sensitive forested watershed in a granite area of subtropical China. We found the precipitation input of inorganic N (17.71 kg N ha⁻¹ year⁻¹ with 54% as NH₄⁺–N) was considerably higher than stream exported inorganic N (5.99 kg N ha⁻¹ year⁻¹ with 83% as NO₃⁻–N), making the watershed a net N sink. The stream export of BCs (151, 1518, 851, and 252 mol ha⁻¹ year⁻¹ for K⁺, Na⁺, Ca²⁺, and Mg²⁺, respectively) was positively correlated (r = 0.80, 0.90, 0.84, and 0.84 for K⁺, Na⁺, Ca²⁺, and Mg²⁺ on a monthly scale, respectively, P < 0.001, n = 36) with PPR_N (389 mol ha⁻¹ year⁻¹) over the three years, suggesting that PPR_N drives loss of BCs in the acid-sensitive ecosystem. A global meta-analysis of 15 watershed studies from non-calcareous ecosystems further supports this hypothesis by showing a similarly strong correlation between ∑BCs output and PPR_N (r = 0.89, P < 0.001, n = 15), in spite of the pronounced differences in environmental settings. Collectively, our results suggest that N transformations rather than anions (NO₃⁻ and/or SO₄²⁻) leaching specifically, are an important mediator of BCs loss in acid-senstive ecosystems. Our study provides the first definitive evidence that the chronic N deposition and subsequent transformation within the watershed drive stream export of BCs through proton production in acid-sensitive ecosystems, irrespective of their current relatively high N retention. Our findings suggest the N-transformation-based proton production can be used as an indicator of watershed outflow quality in the acid-sensitive ecosystems.     

Phenol and sulfide oxidation in a denitrifying biofilm reactor and its microbial community analysis

A microbial consortium attached onto a polyethylene support was used to evaluate the simultaneous oxidation of sulfide and phenol by denitrification. The phenol, sulfide and nitrate loading rates applied to an inverse fluidized bed reactor were up to 168 mg phenol–C/(l d), 37 mg S^2?/(l d) and 168 mg NO₃^?–N/(l d), respectively. Under steady state operation the consumption efficiencies of phenol, sulfide and nitrate were 100%. The N₂ yield (g N₂/g NO₃^?–N) was 0.89. The phenol was mineralized resulting in a yield of 0.82 g bicarbonate–C/g phenol–C and sulfide was completely oxidized to sulfate with a yield of 0.99 g SO₄^2?–S/g S^2?. 16S rRNA gene-based microbial community analysis of the denitrifying biofilm showed the presence of Thauera aromatica, Thiobacillus denitrificans, Thiobacillus sajanensis and Thiobacillus sp. This is the first work reporting the simultaneous oxidation of sulfide and phenol in a denitrifying biofilm reactor.     

Target Binding to S100B Reduces Dynamic Properties and Increases Ca2 +-Binding Affinity for Wild Type and EF-Hand Mutant Proteins

Mutations in the second EF-hand (D61N, D63N, D65N, and E72A) of S100B were used to study its Ca^2 + binding and dynamic properties in the absence and presence of a bound target, TRTK-12. With ^D63NS100B as an exception (^D63NK_D = 50 ± 9 μM), Ca^2 + binding to EF2-hand mutants were reduced by more than 8-fold in the absence of TRTK-12 (^D61NK_D = 412 ± 67 μM, ^D65NK_D = 968 ± 171 μM, and ^E72AK_D = 471 ± 133 μM), when compared to wild-type protein (^WTK_D = 56 ± 9 μM). For the TRTK-12 complexes, the Ca^2 +-binding affinity to wild type (^WT + TRTKK_D = 12 ± 10 μM) and the EF2 mutants was increased by 5- to 14-fold versus in the absence of target (^{D61N + TRTK}K_D = 29 ± 1.2 μM, ^{D63N + TRTK}K_D = 10 ± 2.2 μM, ^{D65N + TRTK}K_D = 73 ± 4.4 μM, and ^{E72A + TRTK}K_D = 18 ± 3.7 μM). In addition, R_ex, as measured using relaxation dispersion for side‐chain ¹⁵N resonances of Asn63 (^D63NS100B), was reduced upon TRTK-12 binding when measured by NMR. Likewise, backbone motions on multiple timescales (picoseconds to milliseconds) throughout wild type, ^D61NS100B, ^D63NS100B, and ^D65NS100B were lowered upon binding TRTK-12. However, the X-ray structures of Ca^2 +-bound (2.0 Å) and TRTK-bound (1.2 Å) ^D63NS100B showed no change in Ca^2 + coordination; thus, these and analogous structural data for the wild-type protein could not be used to explain how target binding increased Ca^2 +-binding affinity in solution. Therefore, a model for how S100B–TRTK‐12 complex formation increases Ca^2 + binding is discussed, which considers changes in protein dynamics upon binding the target TRTK-12.     

Inorganic and organic nitrogen uptake by the toxigenic diatom Pseudo-nitzschia australis (Bacillariophyceae)

The nitrogen uptake capabilities of the toxigenic diatom Pseudo-nitzschia australis (Frenguelli), freshly isolated from Monterey Bay California, were examined in unialgal laboratory cultures at saturating photosynthetic photon flux densities (100 μmol photons m⁻² s⁻¹) and 15 °C. The kinetics of nitrogen (nitrate, ammonium, urea and glutamine) uptake as a function of substrate concentration were estimated from short (20.5 min) incubations using the ¹⁵N-tracer technique. Based on the estimated maximum specific uptake rates and measures of N affinity (the initial slope of the uptake versus nutrient concentration curve), nitrate is the preferred nitrogen substrate, followed by glutamine and ammonium, which are equivalent. Rates of urea uptake by P. australis did not saturate at concentrations as high as 36 μg-at N L⁻¹, and urea uptake as a function of concentration could not be described by Michaelis–Menten kinetics over the concentration gradient tested. Although there is a clear preference for nitrate at equivalent concentrations (compared to ammonium, urea, and glutamine), these laboratory results demonstrate the capability of this pennate diatom to utilize both inorganic and organic forms of nitrogen, supporting field observations that P. australis blooms during both upwelling and non-upwelling conditions off the west coast of North America. Substantial differences in the nitrogenous nutrition of P. australis can be expected in these environments, and anthropogenic inputs of N substrates such as ammonium and urea can support its growth, and may contribute significantly to both harmful diatom blooms and the maintenance of seed populations at non-bloom abundances, particularly during periods of reduced or absent upwelling.     

Nitrate removal and greenhouse gas production in a stream-bed denitrifying bioreactor

Denitrifying bioreactors are currently being tested as an option for treating nitrate (NO₃^?) contamination in groundwater and surface waters. However, a possible side effect of this technology is the production of greenhouse gases (GHG) including nitrous oxide (N₂O) and methane (CH₄). This study examines NO₃^? removal and GHG production in a stream-bed denitrifying bioreactor currently operating in Southern Ontario, Canada. The reactor contains organic carbon material (pine woodchips) intended to promote denitrification. Over a 1 year period, monthly averaged removal of influent (stream water) NO₃^? ranged from 18 to 100% (0.3–2.5 mg N L^?1). Concomitantly, reactor dissolved N₂O and CH₄ production, averaged 6.4 μg N L^?1 (2.4 mg N m^?2 d^?1), and 974 μg C L^?1 (297 mg C m^?2 d^?1) respectively, where production is calculated as the difference between inflow and effluent concentrations. Gas bubbles entrapped in sediments overlying the reactor had a composition ranging from 19 to 64% CH₄, 1 to 6% CO₂, and 0.5 to 2 ppmv N₂O; however, gas bubble emission rates were not quantified in this study. Dissolved N₂O production rates from the bioreactor were similar to emission rates reported for some agricultural croplands (e.g. 0.1–15 mg N m^?2 d^?1) and remained less than the highest rates observed in some N-polluted streams and rivers (e.g. 110 mg N m^?2 d^?1, Grand R., ON). Dissolved N₂O production represented only a small fraction (0.6%) of the observed NO₃^? removal over the monitoring period. Dissolved CH₄ production during summer months (up to 1236 mg C m^?2 d^?1), was higher than reported for some rivers and reservoirs (e.g. 6–66 mg C m^?2 d^?1) but remained lower than rates reported for some wastewater treatment facilities (e.g. sewage treatment plants and constructed wetlands, 19,500–38,000 mg C m^?2 d^?1).     

Nitrate removal processes in a constructed wetland treating drainage from dairy pasture

¹⁵N-labelled NO₃^? was used in a surface-flow constructed wetland in spring to examine the relative importance of competing NO₃^? removal processes. In situ mesocosms (0.25 m²) were dosed with 2 l of ¹⁵NO₃^? (NaNO₃, 300 mg N l^?1, 99 atom% ¹⁵N) and bromide (Br^?) solution (LiBr, 4.3 g l^?1, as a conservative tracer). Concentrations of NO₃^?, Br^?, dissolved oxygen and ¹⁵N₂ were monitored periodically and replicate mesocosms were destructively sampled prior to and 6 days after ¹⁵N addition. Denitrification, immobilisation, plant uptake and dissimilatory NO₃^? reduction to NH₄⁺ (DNRA) accounted for 77, 11, 9 and 2% of ¹⁵NO₃^? transformed during the experiment. Only 6% of denitrification gases were directly measured as atmospheric or dissolved ¹⁵N₂; the remainder (71%) was determined via ¹⁵N mass balance. This indicated that a large proportion of the denitrification gases were entrapped within the soil matrix and/or plant aerenchyma. The floating plant Lemna minor exhibited a significantly higher NO₃^? uptake rate (221 mg kg^?1 d^?1) than Typha orientalis (10 mg kg^?1 d^?1), but periodic harvest of plants would remove <3% of annual NO₃^? inputs. Our results suggest that this 6-year-old constructed wetland functions effectively as a sink for NO₃^? during the growing season with less than one-quarter of the NO₃^? processed sequestered into wetland plant, algal and microbial N pools and the balance permanently removed by denitrification.     

Zooplankton community composition and copepod grazing on the West Florida Shelf in relation to blooms of Karenia brevis

Blooms of the toxin producing dinoflagellate Karenia brevis occur routinely on the West Florida Shelf of the Gulf of Mexico. Nutrient supplies are thought to play a large role in the formation and maintenance of these blooms. The role of top-down control has been less well studied, but grazing, or the lack thereof, on these toxic species may also enhance the formation of large biomass blooms in this region. Zooplankton community structure and copepod species composition were analyzed from samples collected on the West Florida Shelf (WFS) during a NOAA funded ECOHAB regional Karenia Nutrient Dynamics project during October 2007–2010. In 2008 there was no statistical difference in the abundance of zooplankton at bloom and non-bloom stations, however in 2009 there was a statistically significant difference (p < 0.05) between the abundance of zooplankton at stations with Karenia present. To investigate copepod ingestion rates in relation to K. brevis, shipboard and laboratory experiments of the single label method of ¹⁴C labeled phytoplankton culture, and time course ingestion experiments with isolated copepods were performed. Calculated ingestion rates suggest that the copepod species Centropages velificatus, and Acartia tonsa ingested K. brevis, however rates were variable among collection sites and K. brevis strains. Parvocalanus crassirostris did not ingest K. brevis in any of the experiments.     

Comparative diel oxygen cycles preceding and during a Karenia bloom in Sarasota Bay,Florida, USA

The diel change in dissolved oxygen concentrations were recorded with an automated incubator containing a pulsed oxygen sensor in Sarasota Bay, Florida. The deployments occurred during a ‘pre-bloom’ period in May to June 2006, and during a harmful algal bloom dominated by Karenia brevis in September 2006. The diurnal (daylight) increase in dissolved oxygen concentrations varied from 16 to 104 μmol O₂ l⁻¹ with the corresponding nocturnal decrease in oxygen varying from 16 to 77 μmol O₂ l⁻¹. Nocturnal respiration consumed 42–113% of the diurnal net oxygen production with the minimum and maximum during the pre-bloom period. Hourly production rates closely followed fluctuations in irradiance with maximum rates in the late morning. Hourly oxygen utilization rates (community respiration) at night were highest during the first few hours after sunset.     

Structure and magnetic properties of a trinuclear nickel(II) complex with benzenetricarboxylate bridge

Novel trinuclear Ni(II) complex [Ni₃(pmdien)₃(btc)(H₂O)₃](ClO₄)₃ · 4H₂O, 1 where pmdien = N,N,N′,N′,N″-pentamethyldiethylenetriamine, H₃btc = 1,3,5-benzenetricarboxylic (trimesic) acid, has been prepared and structurally characterized. Three nickel atoms are bridged by btc trianion and their coordination sphere is completed by three N atoms of pmdien and O atom of the water molecule. The three nickel(II) magnetic centers are equivalent and their coordination spheres are completed to deformed octahedrons. Magnetic susceptibility was measured over the temperature range 1.8–300 K and zJ′ = ?0.19 cm^?1, D = 3.79 cm^?1, g = 2.18 parameters were calculated.     

Ecosystem functions as indicators for heathland responses to nitrogen fertilisation

Anthropogenic deposition of reactive nitrogen (N) has increased during the 20th century, and is considered an important driver of shifts in ecosystem functions and biodiversity loss. The objective of the present study was to identify those ecosystem functions that best evidence a target ecosystem’s sensitivity to N deposition, taking coastal heathlands as an example. We conducted a three-year field experiment in heathlands of the island Fehmarn (Baltic Sea, North Germany), which currently are subject to a background deposition of 9 kg N ha⁻¹ yr⁻¹. We experimentally applied six levels of N fertilisation (application of 0, 2.5, 5, 10, 20, and 50 kg N ha⁻¹ yr⁻¹), and quantified the growth responses of different plant species of different life forms (dwarf shrubs, graminoids, bryophytes, lichens) as well as shifts in the C:N ratios of plant tissue and humus horizons. For an applicability of the experimental findings (in terms of heathland management and critical load assessment) fertilisation effects on response variables were visualised by calculating the treatment ‘effect sizes’. The current year’s shoot increment of the dominant dwarf shrub Calluna vulgaris proved to be the most sensitive indicator to N fertilisation. Shoot increment significantly responded to additions of ≥ 5 kg N ha⁻¹ yr⁻¹ already in the first year, whereas flower formation of Calluna vulgaris increased only in the high-N treatments. Similarly, tissue C:N ratios of vascular plants (Calluna vulgaris and the graminoids Carex arenaria and Festuca ovina agg.) only decreased in the highest N treatments (50 and 20 kg N ha⁻¹ yr⁻¹, respectively). In contrast, tissue C:N ratios of cryptogams responded more quickly and sensitively than vascular plants. For example, Cladonia spp. tissue C:N ratios responded to N additions ≥ 5 kg N ha⁻¹ yr⁻¹ in the second study year. After three years we observed an increase in cover of graminoids and a corresponding decrease of cryptogams at N fertilisation rates of ≥ 10 kg N ha⁻¹ yr⁻¹. Soil C:N ratios proved to be an inappropriate indicator for N fertilisation at least within our three-year study period. Although current critical N loads for heathlands (10−20 kg N ha⁻¹ yr⁻¹) were confirmed in our experiment, the immediate and highly sensitive response of the current year’s shoots of Calluna vulgaris suggests that at least some ecosystem functions (e.g. dwarf shrub growth) also might respond to low (i.e. < 10 kg N ha⁻¹ yr⁻¹) but chronic inputs of N.     

Availability of N and S affect nutrient acquisition efficiencies differently by Trifolium repens and Lolium perenne when grown in monoculture or in mixture

Sulphur (S) deficiency is recognized as a limiting factor for crop production in many regions in the world. In grasslands, S availability has been shown to alter the biomass production of Trifolium repens and Lolium perenne and their specific interactions. To establish the role of N and S availabilities on the competitive interaction for these minerals by T. repens and L. perenne when grown together, two S rates (0 and 30 kg S ha^?1) combined with three N rates (0, 50 and 180 kg N ha^?1) were investigated in a cut/regrowth experiment over a period of 4 months under glasshouse conditions. N was applied as ¹⁵NH₄ ¹⁵NO₃ to determine their actual N fertilizer recovery in the harvested fraction of the shoot. S yields were used to estimate their apparent S fertilizer recovery. At final harvest, N reserves of T. repens stolons were analyzed to estimate their implication in the regrowth process. In monoculture and in both cuts (1 and 2), N benefited both species by increasing their N and S yields. S benefited only T. repens. In mixture, at cut 1, L. perenne behaved as a better competitor than T. repens thanks to N, while at cut 2, T. repens dominated the community thanks to strong positive S effect. N recovery of L. perenne grown in mixture was greatly improved by S supply. For T. repens, S enhanced its ability to fix N₂ and improved the accumulation of soluble proteins in its stolons. It is clear that the N:S ratio of soil may affect the functionality of grassland plant communities and their structure. Results suggest that (i) the limitations in the availability of soil S could restrict leguminous species growth in high N soil conditions, and (ii) the modulation of S level could be used as a tool to modify the composition of grassland communities.     

Nitrogen uptake kinetics of Prymnesium parvum (Haptophyte)

The uptake rates of different nitrogen (N) forms (NO₃⁻, urea, and the amino acids glycine and glutamic acid) by N-deficient, laboratory-grown cells of the mixotrophic haptophyte, Prymnesium parvum, were measured and the preference by the cells for the different forms determined. Cellular N uptake rates (ρ_cell, fmol N cell⁻¹ h⁻¹) were measured using ¹⁵N-labeled N substrates. P. parvum showed high preference for the tested amino acids, in particular glutamic acid, over urea and NO₃⁻ under the culture nutrient conditions. However, extrapolating these rates to Baltic Seawater summer conditions, P. parvum would be expected to show higher uptake rates of NO₃⁻ and the amino acids relative to urea because of the difference in average concentrations of these substrates. A high uptake rate of glutamic acid at low substrate concentrations suggests that this substrate is likely used through extracellular enzymes. Nitrate, urea and glycine, on the other hand, showed a non-saturating uptake over the tested substrate concentration (1–40 μM-N for NO₃⁻ and urea, 0.5–10 μM-N for glycine), indicating slower membrane-transport rates for these substrates.