Epiphytic lichen indication of nitrogen deposition and climate in the northern rocky mountains,USA

Lichen bioindication can provide economical and spatially extensive monitoring of climate and pollution impacts on ecological communities. We used non-metric multidimensional scaling of lichen community composition and generalized additive models to analyze regional climate and pollution gradients in the northern Rocky Mountains, U.S. Temperature extremes, relative humidity, and N-deposition were strongly related to lichen community composition. Eutrophic species (genera Physcia, Xanthomendoza, and Xanthoria) were associated with high N deposition, low precipitation, and temperature extremes. Estimated N deposition in our study ranged from <0.5 to 4.26 kg N ha⁻¹ year⁻¹ with degradation to lichen communities observed at 4.0 kg N ha⁻¹ year⁻¹, the indicated critical load. The resulting model can track changes in climate and N pollution related to lichen communities over time, identify probable sensitive or impacted habitats, and provide key information for natural resource management and conservation. The approach is broadly applicable to temperate ecosystems worldwide.     

The role of plants in the removal of nutrients at a constructed wetland treating agricultural (dairy) wastewater,Ontario, Canada

The South Nation River Watershed, in eastern Ontario, Canada, is an agricultural watershed impacted by excess nutrient loading primarily from agricultural activities. A constructed wetland for the treatment of agricultural wastewater from a 150-cow dairy operation in this watershed was monitored in its eighth operating season to evaluate the proportion of total nitrogen (TN) (approximated by total Kjeldahl nitrogen (TKN) due to low NO₃⁻) and total phosphorus (TP) removal that could be attributed to storage in Typha latifolia L. and Typha angustifolia L., which dominate this system. Nutrient loading rates were high, with 16.2 kg ha⁻¹ d⁻¹ N and 3.4 kg ha⁻¹ d⁻¹ P entering the wetland and loading the first wetland cell. Plant uptake accounted for 0.7% of TKN removal when the vegetated free water surface cells were considered together. However, separately, in the second wetland cell with lower N and P loading rates, plants accounted for 9% of TKN, 21% of NH₄⁺ and 5% of TP removal. Plant uptake was significant to overall removal given wetland age and nutrient loading. Nutrient storage during the growing season at this constructed wetland helped reduce the nutrient load entering the watershed, already stressed by intensive local agriculture.     

Increasing nitrogen depositions can reduce lichen viability and limit winter food for an endangered Chinese monkey

Increasing economic growth and industrial development in China is starting to impact even remote areas such as the Shennongjia nature reserve, where nitrogen pollution is becoming a major environmental threat. The epiphytic lichen flora is particularly rich in this area and is one of the components of this habitat most sensitive to nitrogen pollution. Since lichens represent an important food resource for the endangered monkey species Rhinopithecus roxellana, a reduction in lichen availability would have harmful consequences for the conservation of its habitat in the Shennongjia Mountains. To investigate the effects of increased nitrogen availability on the local lichen communities, so far scarcely considered, we conducted a one-year field experiment measuring growth, survival, and phosphomonoesterase activity of the widespread species Usnea luridorufa in response to nitrogen (up to 50 kg N ha⁻¹ year⁻¹ deposition) and phosphorus supply. Growth and survival of thalli and propagules of U. luridorufa decreased when treated with N deposition >12.05 kg N ha⁻¹ year⁻¹ and >2.14 kg N ha⁻¹ year⁻¹, respectively. The important role of phosphorus availability in relation to nitrogen supply was demonstrated by the increase in phosphomonoesterase activity with increasing nitrogen availability until a nitrogen toxicity threshold was reached. However, the high concentration of phosphorus in rainwater showed that phosphorus is not a limiting nutrient in the area.The results make a contribution to the knowledge of the negative effects of increased N deposition in the Shennongjia forest ecosystem.     

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.     

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.     

The effects of NH4+ and NO3− on growth,resource allocation and nitrogen uptake kinetics of Phragmites australis and Glyceria maxima

The effects of NH₄⁺ or NO₃⁻ on growth, resource allocation and nitrogen (N) uptake kinetics of two common helophytes Phragmites australis (Cav.) Trin. ex Steudel and Glyceria maxima (Hartm.) Holmb. were studied in semi steady-state hydroponic cultures. At a steady-state nitrogen availability of 34 μM the growth rate of Phragmites was not affected by the N form (mean RGR = 35.4 mg g⁻¹ d⁻¹), whereas the growth rate of Glyceria was 16% higher in NH₄⁺-N cultures than in NO₃⁻-N cultures (mean = 66.7 and 57.4 mg g⁻¹ d⁻¹ of NH₄⁺ and NO₃⁻ treated plants, respectively). Phragmites and Glyceria had higher S/R ratio in NH₄⁺ cultures than in NO₃⁻ cultures, 123.5 and 129.7%, respectively.Species differed in the nitrogen utilisation. In Glyceria, the relative tissue N content was higher than in Phragmites and was increased in NH₄⁺ treated plants by 16%. The tissue NH₄⁺ concentration (mean = 1.6 μmol g fresh wt⁻¹) was not affected by N treatment, whereas NO₃⁻ contents were higher in NO₃⁻ (mean = 1.5 μmol g fresh wt⁻¹) than in NH₄⁺ (mean = 0.4 μmol g fresh wt⁻¹) treated plants. In Phragmites, NH₄⁺ (mean = 1.6 μmol g fresh wt⁻¹) and NO₃⁻ (mean = 0.2 μmol g fresh wt⁻¹) contents were not affected by the N regime. Species did not differ in NH₄⁺ (mean = 56.5 μmol g⁻¹ root dry wt h⁻¹) and NO₃⁻ (mean = 34.5 μmol g⁻¹ root dry wt h⁻¹) maximum uptake rates (V_max), and V_max for NH₄⁺ uptake was not affected by N treatment. The uptake rate of NO₃⁻ was low in NH₄⁺ treated plants, and an induction phase for NO₃⁻ was observed in NH₄⁺ treated Phragmites but not in Glyceria. Phragmites had low K_m (mean = 4.5 μM) and high affinity (10.3 l g⁻¹ root dry wt h⁻¹) for both ions compared to Glyceria (K_m = 6.3 μM, affinity = 8.0 l g⁻¹ root dry wt h⁻¹). The results showed different plasticity of Phragmites and Glyceria toward N source. The positive response to NH₄⁺-N source may participates in the observed success of Glyceria at NH₄⁺ rich sites, although other factors have to be considered. Higher plasticity of Phragmites toward low nutrient availability may favour this species at oligotrophic sites.     

Uptake and inhibition kinetics of nitrogen in Microcystis aeruginosa: Results from cultures and field assemblages collected in the San Francisco Bay Delta,CA

The nitrogen (N) uptake kinetic parameters for Microcystis field assemblages collected from the San Francisco Bay Delta (Delta) in 2012 and non-toxic and toxic laboratory culture strains of M. aeruginosa were assessed. The ¹⁵N tracer technique was used to investigate uptake of ammonium (NH₄⁺), nitrate (NO₃⁻), urea and glutamic acid over short-term incubations (0.5–1 h), and to study inhibition of NO₃⁻, NH₄⁺ and urea uptake by NH₄⁺, NO₃⁻ and NH₄⁺, respectively. This study demonstrates that Delta Microcystis can utilize different forms of inorganic and organic N, with the greatest capacity for NH₄⁺ uptake and the least for glutamic acid uptake, although N uptake did not always follow the classic Michaelis–Menten hyperbolic relationship at substrate concentrations up to 67 μmol N L⁻¹. Current ambient N concentrations in the Delta may be at sub-saturating levels for N uptake, indicating that if N loading (especially NH₄⁺) were to increase, Delta Microcystis assemblages have the potential for increased N uptake rates. Delta Microcystis had the highest specific affinity, α, for NH₄⁺ and the lowest for NO₃⁻. In culture, N uptake by non-toxic and toxic M. aeruginosa strains was much higher than from the field, but followed similar N utilization trends to those in the field. Neither strain showed severe inhibition of NO₃⁻ uptake by NH₄⁺ or inhibition of NH₄⁺ uptake on NO₃⁻, but both strains showed some inhibition of urea uptake by NH₄⁺.     

Effect of nitrogen load on growth and photosynthesis of seedlings of the hybrid larch F1 (Larix gmelinii var. japonica × L. kaempferi) grown on serpentine soil

We studied the growth and photosynthesis of the hybrid larch F₁ (Larix gmelinii var. japonica × L. kaempferi) grown on serpentine soil and the effects of soil N load, to determine the performance of this species as reforestation material in serpentine regions. We prepared 16 experimental plots (2 m × 4 m each), eight on serpentine and eight on brown forest soil, and planted one-year-old cutting seedlings of the hybrid larch F₁ in each plot, in May 2007. Ammonium sulfate was supplied to half of the plots of each soil type in 2008 and 2009, at a load of 47 kg N ha⁻¹ year⁻¹. Although the growth and photosynthetic capacity of hybrid larch F₁ seedlings in the serpentine soil were limited, the rate of growth in serpentine soil was greater than that of Sakhalin spruce (Picea glehnii) that is dominant species in serpentine regions. There was significant interaction between soil type and N load for the growth and photosynthetic parameters. The N load adversely affected growth and photosynthetic parameters in the serpentine soil, while improved them in brown forest soil. Although the growth rate of hybrid larch F₁ without N loading showed high potential as an afforestation species in serpentine region, increasing deposition of N might be a threat to the growth and photosynthesis of the hybrid larch F₁ in serpentine soil.     

Long-term changes of ecosystem services at Solling,Germany: Recovery from acidification,but increasing nitrogen saturation?

Time series of values of ingenious parameters indicating ecosystem services from European beech and Norway spruce ecosystems at Solling, Germany, were evaluated with respect to resilient or adaptive behaviour. Studied indicators comprise the use of monitoring data with up to more than 40 years of observation on deposition of potential acidity, sulphate (SO₄²⁻) budgets, exchangeable base cation pools, Bc/Al ratio in soil solution, nitrogen (N) budgets, foliar nutrition as indicated by the foliar Bc/N ratio, and defoliation. Deposition of potential acidity decreased considerably at both ecosystems. SO₄²⁻ budgets reveal retention of sulphur in the soils affecting acid/base budgets. Exchangeable base cation pools decreased at both ecosystems by about 60%. Bc/Al ratio in soil solution in the mineral soil was mostly below critical limits indicating potential toxic stress to tree roots. N retention in the soils decreased from about 40 kg ha⁻¹ yr⁻¹ in the 1970s to currently very low rates of 0–20 kg ha⁻¹ yr⁻¹ indicating increasing N saturation. Foliar Bc/N ratio decreased at the spruce ecosystem indicating possible nutrient imbalances. Defoliation at both Solling ecosystems is on a high level compared to other forests in Germany, but reveals no distinct relation to soil acidification or N saturation. From the selected indicators, SO₄²⁻ and N budgets reveal resilient behaviour, whereas indicators related to the acid/base status tend to adaptive behaviour.     

Spatiotemporal modeling of watershed nutrient transport dynamics: Implications for eutrophication abatement

The main objective of this study was to quantify nutrient transport dynamics of a previously ungauged, temperate watershed (145 km²) surrounding a shallow eutrophic lake and discern lake response to external nutrient loading, based on soil water assessment tool (SWAT) and the Organization of Economic Cooperation and Development (OECD) empirical lake models, respectively. A SWAT model was used to simulate baseline nutrient dynamics after its calibration and validation against daily tributary flow, total dissolved phosphorus (TDP), total phosphorus (TP), and nitrate (NO₃) loads. On the watershed scale, median annual TDP, TP, and NO₃ losses were 0.4, 1.1, and 2.0 kg ha^− 1, respectively. The highest median annual TP and NO₃ losses were estimated at 3.7 and 7.7 kg ha^− 1 for pastureland and 1.7 and 3.8 kg ha^− 1 for cropland and mixed forests, respectively. Baseflow was the major nutrient transport pathway over a wide range of precipitation events (450 to 900 mm yr^− 1). Erosion was the predominant surface process exporting P across the watershed. Critical source areas (CSAs) of TP and NO₃ comprised 17% and 4% of the watershed, respectively. Annual mean TP, and mean and maximum chlorophyll content indicated a hyper-eutrophication risk for the lake. An external P load reduction by excess of 80% could be necessary to restore mesotrophy in the lake. Our results suggested that subsurface P transport should not be overlooked a priori when groundwater-dependent and extensively farmed watersheds are managed for eutrophication abatement.     

Financial compensation for biodiversity conservation in Ba Be National Park of Northern Vietnam

Terrestrial biodiversity is seriously threatened by an increasing deforestation rate. Financial incentives can stimulate local people to participate in biodiversity conservation. A common approach to derive financial compensation is estimating the opportunity costs arising from the adoption of conservation-friendly land-use practices. Assuming that farmers are risk averse, we used a decision-making under uncertainty approach to derive financial compensation for local people living in and around Ba Be National Park, Vietnam. Moderately risk-averse farmers need US$ 228 ha⁻¹ year⁻¹ financial compensation for the protection of natural forests in the Ecological Rehabilitation and Buffer Zones, whereas the required compensation reaches US$ 334 ha⁻¹ year⁻¹ in the Strict Protection Zone. These compensation payments were derived by comparing financial returns from natural forests with those of the most profitable land-use (maize). Compensation amounts decrease to US$ 130 ha⁻¹ year⁻¹ in the Ecological Rehabilitation and Buffer Zones and to US$ 218 ha⁻¹ year⁻¹ in the Strict Protection Zone, when considering diversification strategy. Derived financial compensations assume that farms comprise 20% croplands and 80% forest. We conclude that compensation amounts vary with farmers’ risk preferences, whether or not diversification strategy is considered, and the level of protection of a given area.     

Patch dynamics of the Mediterranean seagrass Posidonia oceanica: Implications for recolonisation process

Patch dynamics of the Mediterranean slow-growing seagrass Posidonia oceanica was studied in two shallow sites (3–10 m) of the Balearic Archipelago (Spain) through repeated censuses (1–2 year⁻¹). In the sheltered site of Es Port Bay (Cabrera Island), initial patch density (October 2001) was low: 0.05 patches m⁻², and the patch size (number of shoots) distribution was bimodal: most of the patches had less than 6 shoots or between 20 and 50 shoots. Mean patch recruitment in Es Port Bay (0.006 ± 0.002 patches m⁻² year⁻¹) exceeded mean patch loss (0.001 ± 0.001 patches m⁻² year⁻¹), yielding positive net patch recruitment (0.004 ± 0.003 patches m⁻² year⁻¹) and a slightly increased patch density 3 years later (July 2004, 0.06 patches m⁻²). In the exposed site of S’Estanyol, the initial patch density was higher (1.38 patches m⁻², August 2003), and patch size frequency decreased exponentially with size. Patch recruitment (0.26 patches m⁻² year⁻¹) and loss (0.24 patches m⁻² year⁻¹) were high, yielding a slightly increased patch density in the area 1 year later (October 2004, 1.40 patches m⁻²). Most recruited patches consisted of rooting vegetative fragments of 1–2 shoots. Seedling recruitment was observed in Summer 2004 at both sites. Episodic, seedling recruitment comprised 30% and 25% of total patch recruitment in Es Port Bay and S’Estanyol, respectively. Patch survival increased with patch size and no direct removal was observed among patches of 5 shoots or more. Most patches grew along the study, shifting patch distribution towards larger sizes. Within the size range studied (1–150 shoots), absolute shoot recruitment (shoots year⁻¹) increased linearly with patch size (R² = 0.64, p < 4 × 10⁻⁵, N = 125), while specific shoot recruitment was constant (about 0.25 ± 0.05 year⁻¹), although its variance was large for small patches. Given the slow growth rate and the high survival of patches with 5 or more shoots, even the low patch recruitment rates reported here could play a significant role in the colonisation process of P. oceanica.     

Two ways to achieve an anammox influent from real reject water treatment at lab-scale: Partial SBR nitrification and SHARON process

A comparative study to produce the correct influent for Anammox process from anaerobic sludge reject water (700–800 mg NH₄⁺-N L⁻¹) was considered here. The influent for the Anammox process must be composed of NH₄⁺-N and NO₂⁻-N in a ratio 1:1 and therefore only a partial nitrification of ammonium to nitrite is required. The modifications of parameters (temperature, ammonium concentration, pH and solid retention time) allows to achieve this partial nitrification with a final effluent only composed by NH₄⁺-N and NO₂⁻-N at the right stoichiometric ratio. The equal ratio of HCO₃⁻/NH₄⁺ in reject water results in a natural pH decrease when approximately 50% of NH₄⁺ is oxidised. A Sequencing batch reactor (SBR) and a chemostat type of reactor (single-reactor high activity ammonia removal over nitrite (SHARON) process) were studied to obtain the required Anammox influent. At steady state conditions, both systems had a specific conversion rate around 40 mg NH₄⁺-N g⁻¹ volatile suspended solids (VSS) h⁻¹, but in terms of absolute nitrogen removal the SBR conversion was 1.1 kg N day⁻¹ m⁻³, whereas in the SHARON chemostat was 0.35 kg N day⁻¹ m⁻³ due to the different hydraulic retention time (HRT) used. Both systems are compared from operational (including starvation experiments) and kinetic point of view and their advantages/disadvantages are discussed.     

THE EFFECT OF OXYGEN DEPRIVATION ON INORGANIC NITROGEN UPTAKE IN AN ANTARCTIC MACROLICHEN

Snow meltwater containing 36 ng ml⁻¹NO₃⁻-N (raised here to between 95–101 ng ml⁻¹NO₃⁻-N) and 112 ng ml⁻¹NH₄⁺-N was sprayed onto illuminatedUsnea sphacelataat 2°C in a 2-1 capacity transparent perspex chamber force-ventilated with either air or O₂- (and CO₂-) free N₂. The NO₃-concentration in meltwater recirculated through a layer ofU. sphacelatafell toc. 8 ng ml⁻¹after 1·25 h. Although the pattern of decline was broadly comparable in both air and N₂, the initial rate of decline was lower in N₂. When undepleted meltwater was continuously sprayed onto the lichen and the effluent collected for analysis, the lichen was found to retain 55% of the wet deposited NO₃⁻in air but only 27% under N₂. Up to 90% of NH₄⁺supplied in a continuous spray of meltwater was retained by the lichen but this was affected little by O₂and CO₂deprivation.     

Whole-system phosphorus balances as a practical tool for lake management

Controlling phosphorus (P) inputs to lakes remains a priority of lake management. This study develops watershed P balances for 11 recreational lakes in Minnesota. Areal P input rates to the watersheds ranged from 0.32 to 6.0 kg P ha⁻¹ year⁻¹ and was linearly related to the percentage of watershed in agriculture. Watershed P retention ranged from 10% to 89% of input P. Although many best management practices work to increase P retention (by trapping P in basins; reducing erosion; filtration), P retention is not sustainable indefinitely, particularly in “hot spots” such as septic leach fields and heavily manured fields. The watershed P balance tool is a framework that can allow watershed managers to develop novel strategies for managing P. P management strategies should be developed to keep P inputs and exports in balance so that P does not accumulate; long-term P accumulation is not sustainable and can eventually lead to lake eutrophication.     

Urine nitrification with a synthetic microbial community

During long-term extra-terrestrial missions, food is limited and waste is generated. By recycling valuable nutrients from this waste via regenerative life support systems, food can be produced in space. Astronauts’ urine can, for instance, be nitrified by micro-organisms into a liquid nitrate fertilizer for plant growth in space. Due to stringent conditions in space, microbial communities need to be be defined (gnotobiotic); therefore, synthetic rather than mixed microbial communities are preferred. For urine nitrification, synthetic communities face challenges, such as from salinity, ureolysis, and organics.In this study, a synthetic microbial community containing an AOB (Nitrosomonas europaea), NOB (Nitrobacter winogradskyi), and three ureolytic heterotrophs (Pseudomonas fluorescens, Acidovorax delafieldii, and Delftia acidovorans) was compiled and evaluated for these challenges. In reactor 1, salt adaptation of the ammonium-fed AOB and NOB co-culture was possible up to 45 mS cm⁻¹, which resembled undiluted nitrified urine, while maintaining a 44 ± 10 mg NH₄⁺–N L⁻¹ d⁻¹ removal rate. In reactor 2, the nitrifiers and ureolytic heterotrophs were fed with urine and achieved a 15 ± 6 mg NO₃⁻–N L⁻¹ d⁻¹ production rate for 1% and 10% synthetic and fresh real urine, respectively. Batch activity tests with this community using fresh real urine even reached 29 ± 3 mg N L⁻¹ d⁻¹. Organics removal in the reactor (69 ± 15%) should be optimized to generate a nitrate fertilizer for future space applications.     

Anion inhibition profiles of α-, β- and γ-carbonic anhydrases from the pathogenic bacterium Vibrio cholerae

Among the numerous metalloenzymes known to date, carbonic anhydrase (CA, EC 4.2.1.1) was the first zinc containing one, being discovered decades ago. CA is a hydro-lyase, which catalyzes the following hydration–dehydration reaction: CO₂ + H₂O ⇋ HCO₃⁻ + H⁺. Several CA classes are presently known, including the α-, β-, γ-, δ-, ζ- and η-CAs. In prokaryotes, the existence of genes encoding CAs from at least three classes (α-, β- and γ-class) suggests that these enzymes play a key role in the physiology of these organisms. In many bacteria CAs are essential for the life cycle of microbes and their inhibition leads to growth impairment or growth defects of the pathogen. CAs thus started to be investigated in detail in bacteria, fungi and protozoa with the aim to identify antiinfectives with a novel mechanism of action. Here, we investigated the catalytic activity, biochemical properties and anion inhibition profiles of the three CAs from the bacterial pathogen Vibrio cholera, VchCA, VchCAβ and VchCAγ. The three enzymes are efficient catalysts for CO₂ hydration, with k_cat values ranging between (3.4 − 8.23) × 10⁵ s⁻¹ and k_cat/K_M of (4.1 − 7.0) × 10⁷ M⁻¹ s⁻¹. A set of inorganic anions and small molecules was investigated for inhibition of these enzymes. The most potent VchCAγ inhibitors were N,N-diethyldithiocarbamate, sulfamate, sulfamide, phenylboronic acid and phenylarsonic acid, with K_I values ranging between 44 and 91 μM.     

Uptake of dissolved inorganic and organic nitrogen by the benthic toxic dinoflagellate Ostreopsis cf. ovata

Environmental factors that shape dynamics of benthic toxic blooms are largely unknown. In particular, for the toxic dinoflagellate Ostreopsis cf. ovata, the importance of the availability of nutrients and the contribution of the inorganic and organic pools to growth need to be quantified in marine coastal environments. The present study aimed at characterizing N-uptake of dissolved inorganic and organic sources by O. cf. ovata cells, using the ¹⁵N-labelling technique. Experiments were conducted taking into account potential interactions between nutrient uptake systems as well as variations with the diel cycle. Uptake abilities of O. cf. ovata were parameterized for ammonium (NH₄⁺), nitrate (NO₃⁻) and N-urea, from the estimation of kinetic and inhibition parameters. In the range of 0 to 10 μmol N L⁻¹, kinetic curves showed a clear preference pattern following the ranking NH₄⁺ > NO₃⁻ > N-urea, where the preferential uptake of NH₄⁺ relative to NO₃⁻ was accentuated by an inhibitory effect of NH₄⁺ concentration on NO₃⁻ uptake capabilities. Conversely, under high nutrient concentrations, the preference for NH₄⁺ relative to NO₃⁻ was largely reduced, probably because of the existence of a low-affinity high capacity inducible NO₃⁻ uptake system. Ability to take up nutrients in darkness could not be defined as a competitive advantage for O. cf. ovata. Species competitiveness can also be defined from nutrient uptake kinetic parameters. A strong affinity for NH₄⁺ was observed for O. cf. ovata cells that may partly explain the success of this toxic species during the summer season in the Bay of Villefranche-sur-mer (France).     

Investigation on the properties and kinetics of glucose-fed aerobic granular sludge

Aerobic granulation is a process in which suspended biomass aggregate and form discrete well-defined granules in aerobic systems. To investigate the properties and kinetics of aerobic granular sludge, aerobic granules were cultivated with glucose synthetic wastewater in a series of sequencing batch reactors (SBR). The spherical shaped granules were observed on 8th day with the mean diameter of 0.1 mm. With the organic loading rate (OLR) being increased to 4.0 g COD L⁻¹ d⁻¹, aerobic granules grew matured with spherical shape. The size of granules ranged from 1.2 to 1.8 mm, and the corresponding settling velocity of individual granule was 24.2–36.4 m h⁻¹. The oxygen utilization rate (OUR) of mature granules was 41.90 g O₂ kg MLSS⁻¹ h⁻¹, which was two times higher than that of activated sludge (18.32 g O₂ kg MLSS⁻¹ h⁻¹). The experimental data indicated that the substrate utilization and biomass growth kinetics generally followed Monod's kinetics model. The corresponding kinetic coefficients of k (maximum specific substrate utilization rate), K_s (half velocity coefficient), Y (growth yield coefficient) and K_d (decay coefficient) were determined as follows, k_c = 23.65 d⁻¹, K_c = 3367.05 mg L⁻¹, K_N = 0.038 d⁻¹, K_N = 29.65 mg L⁻¹, Y = 0.1927–0.2022 mg MMLS (mg COD)⁻¹ and K_d = 0.00845–0.0135 d⁻¹, respectively. Those properties of aerobic granules made aerobic granules system had a short setup period, high substrate utilization rate and low sludge production.     

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.