Influence of Agapanthus africanus on nitrification in a vertical subsurface flow constructed wetland

The aim of this study is to evaluate the influence of Agapanthus africanus (A. africanus) on nitrification in a vertical subsurface flow constructed wetlands (VSSFs) system. Two lab-scale VSSFs were operated: a) one was planted with A. africanus (vertical flow planted, VFP), and b) the other was unplanted (vertical flow control, VFC). The operation strategy was divided into three phases and consisted of increasing the ammoniacal nitrogen loading rate (ALR) (Phase I: 1.4; Phase II: 2.4; Phase III: 4.4 g NH₄⁺-N·m^?2·d^?1). Nitrification was evaluated in the system at two different depths in the VSSFs (30.5 cm and 60.3 cm, from the top of the system).

The removal efficiencies of COD, BOD₅, TP, and PO₄^?3-P were above 40% in the VFP and VFC during all operation. The mean removal efficiencies of NH₄⁺-N were above 70%. Nitrification was the principal NH₄⁺-N removal mechanism in both systems and transformed more than 50% of the NH₄⁺-N to NO₃^?-N. In terms of the effect of A. africanus on NH₄⁺-N removal during the three operational phases, nonsignificant differences between the two VSSFs were noted (p > 0.05). Thus, A. africanus did not influence nitrification. Finally, the analysis at different depths showed that nitrification occurred in the upper 30.5 cm.     

Greenhouse gas emissions from the energy crop oilseed rape (Brassica napus); the role of photosynthetically active radiation in diurnal N2O flux variation

Oilseed rape (OSR, Brassica napus L.) is an important feedstock for biodiesel; hence, carbon dioxide (CO₂), methane (CH₄) and particularly fertilizer‐derived nitrous oxide (N₂O) emissions during cultivation must be quantified to assess putative greenhouse gas (GHG) savings, thus creating an urgent and increasing need for such data. Substrates of nitrification [ammonium (NH₄)] and denitrification [nitrate (NO₃)], the predominant N₂O production pathways, were supplied separately and in combination to OSR in a UK field trial aiming to: (i) produce an accurate GHG budget of fertilizer application; (ii) characterize short‐ to medium‐term variation in GHG fluxes; (iii) establish the processes driving N₂O emission. Three treatments were applied twice, 1 week apart: ammonium nitrate fertilizer (NH₄NO₃, 69 kg‐N ha^?1) mimicking the farm management, ammonium chloride (NH₄Cl, 34.4 kg‐N ha^?1) and sodium nitrate (NaNO₃, 34.6 kg‐N ha^?1). We deployed SkyLine2D for the very first time, a novel automated chamber system to measure CO_2, CH₄ and N₂O fluxes at unprecedented high temporal and spatial resolution from OSR. During 3 weeks following the fertilizer application, CH₄ fluxes were negligible, but all treatments were a net sink for CO₂ (ca. 100 g CO₂ m^?2). Cumulative N₂O emissions (ca. 120 g CO₂‐eq m^?2) from NH₄NO₃ were significantly greater (P < 0.04) than from NaNO₃ (ca. 80 g CO₂‐eq m^?2), but did not differ from NH₄Cl (ca. 100 g CO₂‐eq m^?2) and reduced the carbon sink of photosynthesis so that OSR was a net GHG source in the fertilizer treatment. Diurnal variation in N₂O emissions, peaking in the afternoon, was more strongly associated with photosynthetically active radiation (PAR) than temperature. This suggests that the supply of carbon (C) from photosynthate may have been the key driver of the observed diurnal pattern in N₂O emission and thus should be considered in future process‐based models of GHG emissions.     

Seasonal and geomorphic controls on N and P removal in riparian zones of the US Midwest

Riparian zones are an important strategy to mitigate N and P export to streams. However, their efficiency with respect to nitrate (NO₃ ^?), ammonium (NH₄ ⁺), or soluble reactive phosphorus (SRP) in groundwater remains uncertain in the US Midwest. This study investigates water table fluctuations and NO₃ ^?, NH₄ ⁺, and SRP concentration dynamics in two riparian zone types (outwash vs. glacial till) common in the upper US Midwest. During low water table periods, NO₃ ^? removal was 93 % at WR (outwash site), and 75 % at LWD (glacial till site); but during high water table periods, NO₃ ^? removal efficiencies dropped to 50 % at WR, and 14 % at LWD. Median seasonal mass fluxes of NO₃ ^? removed at WR (9.4–21.7 mg N day^?1 m^?1 of stream length) and LWD (0.4–1.9 mg N day^?1 m^?1) were small compared to other riparian zones in glaciated landscapes. The WR site was a small SRP sink (0.114 and 0.118 mg day^?1 m^?1 during the dry period and wet period, respectively), while LWD acted as a small SRP source to the stream (0.004 mg day^?1 m^?1 during the dry period; 0.075 mg day^?1 m^?1 during the wet period). Both LWD and WR acted as sources of NH₄ ⁺ to the stream with mass fluxes ranging from 0.17 to 7.75 mg N day^?1 m^?1. Although riparian zones in the US Midwest provide many ecosystem services, results suggest they are unlikely to efficiently mitigate N and P pollution in subsurface flow.     

Kinetic Modeling for Simultaneous Organic Carbon Oxidation,Nitrification, and Denitrification of Abattoir Wastewater in Sequencing Batch Reactor

A laboratory-scale study was conducted in a 20.0-L sequencing batch reactor (SBR) to explore the feasibility of simultaneous removal of organic carbon and nitrogen from abattoir wastewater. The reactor was operated under three different combinations of aerobic-anoxic sequence, viz., (4+4), (5+3), and (5+4) h of total react period, with influent soluble chemical oxygen demand (SCOD) and ammonia nitrogen (NH₄⁺-N) level of 2200 ± 50 and 125 ± 5 mg L^?1, respectively. In (5+4) h cycle, a maximum 90.27% of ammonia reduction corresponding to initial NH₄⁺-N value of 122.25 mg L^?1 and 91.36% of organic carbon removal corresponding to initial SCOD value of 2215.25 mg L^?1 have been achieved, respectively. The biokinetic parameters such as yield coefficient (Y), endogenous decay constant (k_d), and half-velocity constant (K_s) were also determined to improve the design and operation of package effluent treatment plants comprising SBR units. The specific denitrification rate (q_DN) during anoxic condition was estimated as 6.135 mg N/g mixed liquor volatile suspended solid (MLVSS)·h on 4-h average contact period. The value of Y, k_d and K_s for carbon oxidation and nitrification were found to be in the range of 0.6225–0.6952 mg VSS/mg SCOD, 0.0481–0.0588 day^?1, and 306.56–320.51 mg L^?1, and 0.2461–0.2541 mg VSS/mg NH₄⁺-N, 0.0324–0.0565 day^?1, and 38.28–50.08 mg L^?1, respectively, for different combinations of react periods.     

Nitrogen and phosphorus removal by wetland mesocosms subjected to different hydroperiods

The effect of hydroperiod on nutrient removal efficiency from simulated wastewater was investigated in replicate wetland mesocosms (area, 2 m², planted with Scirpus californicus). Alternate draining and flooding of sediments (pulsed discharge) increased nutrient removal efficiency compared to the continuous-flow “control”. Average PO₄³⁻ removal efficiency was 20–30% higher in wetland mesocosms that drained twice daily compared to the control. Inorganic N removal efficiency was less affected than phosphate removal by hydroperiod variation. At the higher NH₄⁺ loading rate (1.83 g N m⁻² day⁻¹), inorganic N removal efficiency was consistently 5–20% higher in pulsed-discharge wetland mesocosms than in the control. At the lower NH₄⁺ loading rate (0.9 g N m ⁻² day ⁻¹), pulsed-discharge hydrology had no effect on inorganic N removal efficiency. Twice-daily drainage exhibited average inorganic N removal efficiencies of 96% (lower N loading rate) and 87% (higher N loading) and average phosphate removal efficiencies of 81% (lower P loading) and 90% (higher P loading). Mass balance data from the continuous-flow treatment revealed that the aquatic macrophyte Scirpus californicus was the most important nutrient sink, assimilating 50% of the NH₄⁺ and PO₄³⁻ supply. The high plant productivity in the mesocosms (15.6 kg m⁻² year⁻¹) occurred under conditions of high light (high edge per mesocosm area) and high root contact with nutrient-rich influent (shallow, sandy substrate) and may overestimate plant uptake in larger wetlands. The addition of a nitrification-inhibitor (N-Serve) indicated that 34% of the NH₄⁺ supply was transformed to NO₃⁻ by nitrifying bacteria.     

闽江河口湿地土壤CH4产生与氧化速率对外源氮、硫添加的响应

通过室内培养实验,研究了外源氮、硫添加对闽江河口湿地土壤CH_4产生/氧化速率以及土壤理化性质的短期影响。NH_4Cl(N1)和NH_4NO_3(N3)处理在各培养阶段均显著促进土壤CH_4产生速率(P0.05),较对照分别提高136.70%和136.55%;NH_4Cl+K_2SO_4(NS1)和NH_4NO_3+K_2SO_4(NS3)处理在培养第3、6、12、15和18天均显著促进了CH_4产生速率(P0.05)。KNO_3(N2)、K_2SO_4(S)处理在不同培养时间对CH_4产生速率影响均不显著(P0.05);KNO_3+K_2SO_4(NS2)处理除在第21天外(P0.05),其他时间影响均不显著(P0.05)。N2、N3、NS2和NS3处理均显著促进了土壤CH_4氧化速率(P0.05),平均CH4氧化速率较CK分别提高了145.30%、142.93%、139.48%和112.68%。整体而言,不同添加处理并没有显著改变湿地土壤CH_4产生/氧化速率的时间变化规律,各处理均表现为随培养时间先增加而后逐渐降低。短期培养结束后,土壤可溶性有机碳(DOC)、电导率、p H值在不同处理间均不存在显著差异(P0.05);土壤NH+4-N含量在N1、N3、NS1和NS3处理下,NO_3~--N含量在N2、N3、NS2和NS3处理下,SO_4~(2-)含量在S、NS1、NS2和NS3处理下均显著高于对照处理(P0.05)。相关分析显示,DOC、铵态氮(NH+4-N)和硝态氮(NO_3~--N)是氮、硫添加处理下影响闽江河口湿地土壤CH_4产生/氧化速率短期变化的主要控制因素。     

NH4 + transport system of a psychrophilic marine bacterium, Vibrio sp. strain ABE-1

NH₄ ⁺ transport system of a psychrophilic marine bacterium Vibrio sp. strain ABE-1 (Vibrio ABE-1) was examined by measuring the uptake of [¹⁴C]methylammonium ion (¹⁴CH₃NH^{3 +}) into the intact cells. ¹⁴CH₃NH₃ ⁺ uptake was detected in cells grown in medium containing glutamate as the sole nitrogen source, but not in those grown in medium containing NH₄Cl instead of glutamate. Vibrio ABE-1 did not utilize CH₃NH₃ ⁺ as a carbon or nitrogen source. NH₄Cl and nonradiolabeled CH₃NH₃ ⁺ completely inhibited ¹⁴CH₃NH₃ ⁺ uptake. These results indicate that ¹⁴CH₃NH₃ ⁺ uptake in this bacterium is mediated via an NH₄ ⁺ transport system and not by a specific carrier for CH₃NH₃ ⁺. The respiratory substrate succinate was required to drive ¹⁴CH₃NH₃ ⁺ uptake and the uptake was completely inhibited by KCN, indicating that the uptake was energy dependent. The electrochemical potentials of H⁺ and/or Na⁺ across membranes were suggested to be the driving forces for the transport system because the ionophores carbonylcyanide m-chlorophenylhydrazone and monensin strongly inhibited uptake activities at pH 6.5 and 8.5, respectively. Furthermore, KCl activated ¹⁴CH₃NH₃ ⁺ uptake. The ¹⁴CH₃NH₃ ⁺ uptake activity of Vibrio ABE-1 was markedly high at temperatures between 0° and 15°C, and the apparent K _m value for CH₃NH₃ ⁺ of the uptake did not change significantly over the temperature range from 0° to 25°C. Thus, the NH₄ ⁺ transport system of this bacterium was highly active at low temperatures. Received: August 1, 1998 / Accepted: October 8, 1998     

Nitrogen Acquisition from Atmospheric NH3 by Lolium perenne

Ammonia (NH₃) is the third most abundant N species in the atmosphere and, due to various natural and anthropogenic sources, can reach high concentrations in some areas. While some plants show effects of toxicity, others are capable of using this N-form and grow well without any utilization of soil-N. Acquisition of atmospheric NH₃ will affect the acid-base balance of the plants as absorption and dissolution causes an alkalinisation (production of OH^?) and assimilation of NH₃ results in an acidification (generation of H⁺). As there is only a limited capacity for biochemical disposal of excess H⁺ in shoots, pH regulation may involve H⁺/OH^? extrusion into the media via roots and transport of (in)organic ions between roots and above-ground parts of the plant. Our aim therefore was to assess NH₃ acquisition by Lolium perenne and to study the effects of gas phase NH₃ on growth, acid-base balance and mineral composition of the plants. The experiments therefore included application of a range of ¹⁴NH₃ to the shoots and of ¹⁵N as NO₃^?, NH₄⁺ or NH₄NO₃ to the roots, from which the amount of gas phase NH₃ acquisition could be quantified. Analysis of the mineral composition provided data for calculation of acid-base balance as well as for water use efficiencies of the plants. The results indicate that over the range of NH₃ supplied, plants from all treatments could utilize gas-phase NH₃ as demonstrated by increases in growth and in N and C use efficiencies. Plants receiving NO₃^? via their roots had a higher capacity to use gaseous NH₃ than those growing with NH₄⁺. NH₃ assimilation in shoots reduced both the acid load with NH₄⁺ nutrition and the alkaline load with NO₃^? supply to the roots. The results of the experiments are discussed in relation to possible acid-base regulation mechanisms of the whole plant.     

Studies in nitrification of municipal sewage in an upflow biofilter

The upflow aerated biofilter with polyurethane foam cubes as the supporting medium was used for the investigation of nitrification studies on municipal sewage (secondary treated as well as untreated domestic sewage). In case of secondary treated sewage effluent, a synthetic composition of NH₄ ⁺-N and COD of each 50?mg/l was studied for a HRT variation of 24, 12, 8 and 6 hours. The ammonium removal efficiencies were found to be in the range of 98 to 100% with the steady-state effluent concentrations of NH₄ ⁺-N and NO₂ ^?-N in the range of 1–4 mg/l and 0.1–0.2?mg/l respectively. In case of domestic sewage system, nitrification studies along with suspended solids removal study was carried-out on untreated sewage for a HRT variation of 24, 12 and 6 hours. The ammonium removal efficiencies of 100% were observed for all the three HRT values at very high COD/NH₄ ⁺-N ratio of 15. The suspended solids removal efficiencies of 95 to 98% were observed with the average effluent suspended solids concentration of 5.9 to 15.9?mg/l. The experiments were conducted in non-backwash conditions of the biofilter. The study has revealed the best use of the upflow biofilter system for nitrification applications and suspended solids removal.     

Methane efflux in relation to plant biomass and sediment characteristics in stands of three common emergent macrophytes in boreal mesoeutrophic lakes

Methane efflux was studied in stands of three emergent macrophyte species (Equisetum fluviatile, Schoenoplectus lacustris and Phragmites australis) commonly found in the littoral zone of boreal lakes. In vegetation stands with relatively low methane (CH₄) emissions (<0.3 mol m^?2 (ice‐free period)^?1), the seasonal variation of CH₄ efflux was better correlated with the dynamics of plant growth than variation in sediment temperature. In dense and productive vegetation stands that released high amounts of CH₄ (2.3–7.7 mol m^?2 (ice‐free period)^?1), the seasonal variation in CH₄ efflux was correlated with sediment temperature, indicating that methanogens were more limited by temperature than substrate supply. The bottom type at the growth site of the emergent plants significantly influenced the ratio of CH₄ efflux to aboveground biomass of plants (Eff : B). The lowest Eff : B ratio was found in E. fluviatile stands growing on sand bottom under experimental conditions and the highest in P. australis‐dominated littoral areas accumulating detritus from external sources. The future changes expected in the hydrology of boreal lakes and rivers because of climatic warming may impact the growth conditions of aquatic macrophytes as well as decomposition and accumulation of detritus and, thus, CH₄ effluxes from boreal lakes.     

Ammonium and Methylammonium Transport in Egeria densa Leaves in Conditions of Different H+ Pump Activity

Previous data in Egeria densa leaves demonstrated a strong inhibitory effect of Cs⁺ on passive K⁺ influx and on K⁺-induced, ATP-dependent electrogenic proton extrusion. In this paper we analyzed, using the same material, the effects of Cs⁺ on ammonium (NH₄⁺) and methylammonium (CH₃NH₃⁺) transport in order to elucidate whether a common transport system for K⁺ and NH₄⁺ could be demonstrated. The effects of Cs⁺ on NH₄⁺- and CH₃NH₃⁺-induced titratable H⁺ extrusion (–ΔH⁺) and on transmembrane electrical potential difference (E_m) in E. densa leaves were analyzed in parallel. All experiments were run either in the absence or presence of fusicoccin, corresponding to low or high H⁺-ATPase activity and membrane hyperpolarization and leading, in this material, to respectively active or passive transport of K⁺. The results suggest the presence in E. densa leaves of two distinct pathways for NH₄⁺ uptake: one in common with NH₄⁺ and (with lower affinity) CH₃NH₃⁺, insensitive to Cs⁺, and a second system, operating at higher H⁺-ATPase activity and E_m hyperpolarization, strongly inhibited by Cs⁺ and impermeable to CH₃NH₃⁺. In agreement with this hypothesis, Xenopus laevis oocytes injected with the KAT1 RNA of Arabidopsis thaliana were permeable to K⁺ and NH₄⁺, but not to CH₃NH₃⁺.     

Atmospheric methane uptake by tropical montane forest soils and the contribution of organic layers

Microbial oxidation in aerobic soils is the primary biotic sink for atmospheric methane (CH₄), a powerful greenhouse gas. Although tropical forest soils are estimated to globally account for about 28% of annual soil CH₄ consumption (6.2 Tg CH₄ year^?1), limited data are available on CH₄ exchange from tropical montane forests. We present the results of an extensive study on CH₄ exchange from tropical montane forest soils along an elevation gradient (1,000, 2,000, 3,000 m) at different topographic positions (lower slope, mid-slope, ridge position) in southern Ecuador. All soils were net atmospheric CH₄ sinks, with decreasing annual uptake rates from 5.9 kg CH₄–C ha^?1 year^?1 at 1,000 m to 0.6 kg CH₄–C ha^?1 year^?1 at 3,000 m. Topography had no effect on soil atmospheric CH₄ uptake. We detected some unexpected factors controlling net methane fluxes: positive correlations between CH₄ uptake rates, mineral nitrogen content of the mineral soil and with CO₂ emissions indicated that the largest CH₄ uptake corresponded with favorable conditions for microbial activity. Furthermore, we found indications that CH₄ uptake was N limited instead of inhibited by NH₄ ⁺. Finally, we showed that in contrast to temperate regions, substantial high affinity methane oxidation occurred in the thick organic layers which can influence the CH₄ budget of these tropical montane forest soils. Inclusion of elevation as a co-variable will improve regional estimates of methane exchange in these tropical montane forests.     

Kinetics of nitrate and ammonium absorption and accompanying H+ fluxes in roots of Lolium perenne L. and N2-fixing Trifolium repens L.

Kinetic parameters for NH₄⁺ and NO₃^? uptake were measured in intact roots of Lolium perenne and actively N₂-fixing Trifolium repens. Simultaneously, net H⁺ fluxes between the roots and the root medium were recorded, as were the net photosynthetic rate and transpiration of the leaves. A Michaelis–Menten-type high-affinity system operated in the concentration range up to about 500 mmol m^?3 NO₃^? or NH₄⁺. In L. perenne, the V_max of this system was 9–11 and 13–14 μmol g^?1 root FW h^?1 for NO₃^? and NH₄⁺, respectively. The corresponding values in T. repens were 5–7 and 2 μmol g^?1 root FW h^?1. The K_m for NH₄⁺ uptake was much lower in L. perenne than in T. repens (c. 40 compared with 170 mmol m^?3), while K_m values for NO₃^? absorption were roughly similar (around 130 mmol m^?3) in the two species. There were no indications of a significant efflux component in the net uptake of the two ions. The translocation rate to the shoots of nitrogen derived from absorbed NO₃^?-N was higher in T. repens than in L. perenne, while the opposite was the case for nitrogen absorbed as NH₄⁺. Trifolium repens had higher rates of transpiration and net photosynthesis than L. perenne. Measurements of net H⁺ fluxes between roots and nutrient solution showed that L. perenne absorbing NO₃^? had a net uptake of H⁺, while L. perenne with access to NH₄⁺ and T. repens, with access to NO₃^? or NH₄⁺, in all cases acidified the nutrient solution. Within the individual combinations of plant species and inorganic N form, the net H⁺ fluxes varied only a little with external N concentration and, hence, with the absorption rate of inorganic N. Based on assessment of the net H⁺ fluxes in T. repens, nitrogen absorption rate via N₂ fixation was similar to that of inorganic N and was not down-regulated by exposure to inorganic N for 2 h. It is concluded that L. perenne will have a competitive advantage over T. repens with respect to inorganic N acquisition.     

Biotic and abiotic controls on the ecosystem significance of consumer excretion in two contrasting tropical streams

1. Excretion of nitrogen (N) and phosphorus (P) is a direct and potentially important role for aquatic consumers in nutrient cycling that has recently garnered increased attention. The ecosystem‐level significance of excreted nutrients depends on a suite of abiotic and biotic factors, however, and few studies have coupled measurements of excretion with consideration of its likely importance for whole‐system nutrient fluxes. 2. We measured rates and ratios of N and P excretion by shrimps (Xiphocaris elongata and Atya spp.) in two tropical streams that differed strongly in shrimp biomass because a waterfall excluded predatory fish from one site. We also made measurements of shrimp and basal resource carbon (C), N and P content and estimated shrimp densities and ecosystem‐level N and P excretion and uptake. Finally, we used a 3‐year record of discharge and NH₄‐N concentration in the high‐biomass stream to estimate temporal variation in the distance required for excretion to turn over the ambient NH₄‐N pool. 3. Per cent C, N, and P body content of Xiphocaris was significantly higher than that of Atya. Only per cent P body content showed significant negative relationships with body mass. C:N of Atya increased significantly with body mass and was higher than that of Xiphocaris. N : P of Xiphocaris was significantly higher than that of Atya. 4. Excretion rates ranged from 0.16–3.80 μmol NH₄‐N shrimp^?1 h^?1, 0.23–5.76 μmol total dissolved nitrogen (TDN) shrimp^?1 h^?1 and 0.002–0.186 μmol total dissolved phosphorus (TDP) shrimp^?1 h^?1. Body size was generally a strong predictor of excretion rates in both taxa, differing between Xiphocaris and Atya for TDP but not NH₄‐N and TDN. Excretion rates showed statistically significant but weak relationships with body content stoichiometry. 5. Large between‐stream differences in shrimp biomass drove differences in total excretion by the two shrimp communities (22.3 versus 0.20 μmol NH₄‐N m^?2 h^?1, 37.5 versus 0.26 μmol TDN m^?2 h^?1 and 1.1 versus 0.015 μmol TDP m^?2 h^?1), equivalent to 21% and 0.5% of NH₄‐N uptake and 5% and <0.1% of P uptake measured in the high‐ and low‐biomass stream, respectively. Distances required for excretion to turn over the ambient NH₄‐N pool varied more than a hundredfold over the 3‐year record in the high‐shrimp stream, driven by variability in discharge and NH₄‐N concentration. 6. Our results underscore the importance of both biotic and abiotic factors in controlling consumer excretion and its significance for nutrient cycling in aquatic ecosystems. Differences in community‐level excretion rates were related to spatial patterns in shrimp biomass dictated by geomorphology and the presence of predators. Abiotic factors also had important effects through temporal patterns in discharge and nutrient concentrations. Future excretion studies that focus on nutrient cycling should consider both biotic and abiotic factors in assessing the significance of consumer excretion in aquatic ecosystems.     

Microbial enzyme activity,nutrient uptake and nutrient limitation in forested streams

1. We measured NH₄⁺ and PO₄^?3 uptake length (S_w), uptake velocity (V_f), uptake rate (U), biofilm respiration and enzyme activity and channel geomorphology in streams draining forested catchments in the northwestern (Northern California Coast Range and Cascade Mountains) and southeastern (Appalachian and Ouachita mountains) regions of the United States. Our goal was to use measures of biofilm enzyme activity and nutrient uptake to assess nutrient limitation in forested streams across broad regional scales. 2. Geomorphological attributes, biofilm enzyme activity and NH₄⁺ uptake were significantly different among streams in the four study units. There was no study unit effect on PO₄^?3 uptake. The proportion of the stream channel in pools, % woody debris, % canopy closure, median substrate size (d₅₀), stream width (w), stream velocity (v), discharge (Q), dispersion coefficient (D) and transient storage (A_s/A) were correlated with biofilm enzyme activity and nutrient uptake in some study units. 3. Canonical correlation analyses across study units revealed significant correlations of NH₄‐V_f and PO₄‐V_f with geomorphological attributes (w, d₅₀, D, % woody debris, channel slope and % pools) and biofilm phosphatase activity. 4. The results did not support our expectation that carbon processing rates by biofilm microbial assemblages would be governed by stream nutrient availability or that resulting biofilm enzyme activity would be an indicator of nutrient uptake. However, the relative abundances of peptidases, phosphatase and glycosidases did yield insight into potential N‐, P‐ and C‐limitation of stream biofilm assemblages, and our use of biofilm enzyme activity represents a novel application for understanding nutrient limitations in forested streams. 5. Regressions of V_f and U against ambient NH₄⁺ and PO₄^?3 indicated that none of our study streams was either NH₄⁺ or PO₄^?3 saturated. The Appalachian, Ouachita and Coastal streams showed evidence of NH₄⁺ limitation; the Ouachita and Coastal streams were PO₄^?3 limited. As a correlate of nutrient limitation and saturation in streams, ratios of total aminopeptidase and phosphatase activities and the ratio of NH₄‐U to PO₄‐U indicate these forested streams are predominantly N‐limited, with only the streams draining Ouachita and Coastal catchments demonstrating appreciable levels of P‐limitation. 6. Our results comparing the stoichiometry of microbial enzyme activity with nutrient uptake ratios and with the molar ratios N and P in stream waters suggest that biological limitations are not strictly the result of stream chemistry and that the assessments of nutrient limitations in stream ecosystems should not be based on chemistry alone. 7. Our present study, along with previous work in streams, rivers and wetlands, suggests that microbial enzyme activities, especially the ratios of total peptidases to phosphatase, are useful indicators of nutrient limitations in aquatic ecosystems.     

Dry deposition of ammonia gas drives species change faster than wet deposition of ammonium ions: evidence from a long‐term field manipulation

Although the effects of atmospheric nitrogen deposition on species composition are relatively well known, the roles of the different forms of nitrogen, in particular gaseous ammonia (NH₃), have not been tested in the field. Since 2002, we have manipulated the form of N deposition to an ombrotrophic bog, Whim, on deep peat in southern Scotland, with low ambient N (wet + dry = 8 kg N ha^?1 yr^?1) and S (4 kg S ha^?1 yr^?1) deposition. A gradient of ammonia (NH₃, dry N), from 70 kg N ha^?1 yr^?1 down to background, 3–4 kg N ha^?1 yr^?1 was generated by free air release. Wet ammonium (NH₄⁺, wet N) was provided to replicate plots in a fine rainwater spray (NH₄Cl at +8, +24, +56 kg N ha^?1 yr^?1). Automated treatments are coupled to meteorological conditions, in a globally unique, field experiment. Ammonia concentrations were converted to NH₃‐N deposition (kg N ha^?1) using a site/vegetation specific parameterization. Within 3 years, exposure to relatively modest deposition of NH₃, 20–56 kg NH₃‐N ha^?1 yr^?1 led to dramatic reductions in species cover, with almost total loss of Calluna vulgaris, Sphagnum capillifolium and Cladonia portentosa. These effects appear to result from direct foliar uptake and interaction with abiotic and biotic stresses, rather than via effects on the soil. Additional wet N by contrast, significantly increased Calluna cover after 5 years at the 56 kg N dose, but reduced cover of Sphagnum and Cladonia. Cover reductions caused by wet N were significantly different from and much smaller than those caused by equivalent dry N doses. The effects of gaseous NH₃ described here, highlight the potential for ammonia to destroy acid heathland and peat bog ecosystems. Separating the effects of gaseous ammonia and wet ammonium deposition, for a peat bog, has significant implications for regulatory bodies and conservation agencies.     

Use of chemical ionization for GC–MS metabolite profiling

Metabolite profiling is commonly performed by GC–MS of methoximated trimethylsilyl derivatives. The popularity of this technique owes much to the robust, library searchable spectra produced by electron ionization (EI). However, due to extensive fragmentation, EI spectra of trimethylsilyl derivatives are commonly dominated by trimethylsilyl fragments (e.g. m/z 73 and 147) and higher m/z fragment ions with structural information are at low abundance. Consequently different metabolites can have similar EI spectra, and this presents problems for identification of “unknowns” and the detection and deconvolution of overlapping peaks. The aim of this work is to explore use of positive chemical ionization (CI) as an adjunct to EI for GC–MS metabolite profiling. Two reagent gases differing in proton affinity (CH₄ and NH₃) were used to analyse 111 metabolite standards and extracts from plant samples. NH₃-CI mass spectra were simple and generally dominated by [MH]⁺ and/or the adduct [M+NH₄]⁺. For the 111 metabolite standards, m/z 73 and 147 were less than 3% of basepeak in NH₃-CI and less than 30% of basepeak in CH₄-CI. With CH₄-CI, [MH]⁺ was generally present but at lower relative abundance than for NH₃-CI. CH₄-CI spectra were commonly dominated by losses of CH₄ [M+1-16]⁺, 1–3 TMSOH [M+1-nx90]⁺, and combinations of CH₄ and TMSOH losses [M+1-nx90-16]⁺. CH₄-CI and NH₃-CI mass spectra are presented for 111 common metabolites, and CI is used with real samples to help identify overlapping peaks and aid identification via determination of the pseudomolecular ion with NH₃-CI and structural information with CH₄-CI.

     

Further Analysis of Anammox Bacterial Community Structures Along an Anthropogenic Nitrogen-Input Gradient from the Riparian Sediments of the Pearl River Delta to the Deep-Ocean Sediments of the South China Sea

The community structures of anammox bacteria in sediments along an anthropogenic inorganic nitrogen input gradient were further delineated with the newly available information incorporated. Anammox bacterial 16S rRNA gene-amplified sequences retrieved from riparian sediments of the Pearl River, Mai Po coastal wetland, and the South China Sea (SCS) sediments were compiled, compared and analyzed. Results indicated that the community structures of anammox bacteria varied from the upstream of the Pearl River to deep-ocean sediment of the SCS along the anthropogenic input grandient. Mai Po wetland had the most diverse anammox bacteria, followed by the shallow SCS, deep SCS and the Pearl River. Genera of the anammox bacteria Kuenenia and Brocadia showed higher proportion in the riparian sediments of the Pearl River, while those of Kuenenia and Scalindua dominated the Mai Po coastal wetland. The Scalindua subclusters showed apparent segregation in coastal wetland (S. zhenghei-III and S. wagneri), shallow SCS (S. zhenghei-I and S3) and deep SCS (S. zhenghei-I, S2 and S. arabica). Pearson correlation analysis indicated nitrogen species [NH₄⁺ and ∑(NO₂^?+NO₃^? )] negatively correlated with the diversity indices of anammox bacteria. Canonical correspondence analysis (CCA) showed that salinity, inorganic nitrogen [NH₄+, ∑(NO₂^?+NO₃^?)], and ratio of NH₄⁺/∑(NO₂^? +NO₃^?) significantly affected the bacterial community compositions. Results collectively support that the community composition of anammox bacteria can serve as a bio-indicator to the anthropogenic terrestrial N input or pollution.     

Effect of influent substrate ratio on anammox granular sludge: performance and kinetics

Effect of influent substrate ratio on anammox process was studied in sequencing batch reactor. Operating temperature was fixed at 35 ± 1 °C. Influent pH and hydraulic retention time were 7.5 and 6 h, respectively. When influent NO₂ ^?-N/NH₄ ⁺-N was no more than 2.0, total nitrogen removal rate (TNRR) increased whereas NH₄ ⁺-N removal rate stabilized at 0.32 kg/(m³ d). ΔNO₂ ^?-N/ΔNH₄ ⁺-N increased with enhancing NO₂ ^?-N/NH₄ ⁺-N. When NO₂ ^?-N/NH₄ ⁺-N was 4.5, ΔNO₂ ^?-N/ΔNH₄ ⁺-N was 1.98, which was much higher than theoretical value (1.32). The IC₅₀ of NO₂ ^?-N was 289 mg/L and anammox activity was inhibited at high NO₂ ^?-N/NH₄ ⁺-N ratio. With regard to influent NH₄ ⁺-N/NO₂ ^?-N, the maximum NH₄ ⁺-N removal rate was 0.36 kg/(m³ d), which occurred at the ratio of 4.0. Anammox activity was inhibited when influent NH₄ ⁺-N/NO₂ ^?-N was higher than 5.0. With influent NO₃ ^?-N/NH₄ ⁺-N of 2.5–6.5, NH₄ ⁺-N removal rate and NRR were stabilized at 0.33 and 0.40 kg/(m³ d), respectively. When the ratio was higher than 6.5, nitrogen removal would be worsened. The inhibitory threshold concentration of NO₂ ^?-N was lower than NH₄ ⁺-N and NO₃ ^?-N. Anammox bacteria were more sensitive to NO₂ ^?-N than NH₄ ⁺-N and NO₃ ^?-N. TNRR would be enhanced with increasing nitrogen loading rate, but sludge floatation occurred at high nitrogen loading shock. The Han-Levenspiel could be applied to simulate nitrogen removal resulting from NO₂ ^?-N inhibition.     

Identification of functionally active aerobic methanotrophs in sediments from an arctic lake using stable isotope probing

Arctic lakes are a significant source of the greenhouse gas methane (CH₄), but the role that methane oxidizing bacteria (methanotrophs) play in limiting the overall CH₄ flux is poorly understood. Here, we used stable isotope probing (SIP) techniques to identify the metabolically active aerobic methanotrophs in upper sediments (0–1 cm) from an arctic lake in northern Alaska sampled during ice‐free summer conditions. The highest CH₄ oxidation potential was observed in the upper sediment (0–1 cm depth) with 1.59 µmol g wet weight^?1 day^?1 compared with the deeper sediment samples (1–3 cm, 3–5 cm and 5–10 cm), which exhibited CH₄ oxidation potentials below 0.4 µmol g wet weight^?1 day^?1. Both type I and type II methanotrophs were directly detected in the upper sediment total communities using targeted primer sets based on 16S rRNA genes. Sequencing of 16S rRNA genes and functional genes (pmoA and mxaF) in the ¹³C‐DNA from the upper sediment indicated that type I methanotrophs, mainly Methylobacter, Methylosoma, Methylomonas and Methylovulum miyakonense, dominated the assimilation of CH₄. Methylotrophs, including the genera Methylophilus and/or Methylotenera, were also abundant in the ¹³C‐DNA. Our results show that a diverse microbial consortium acquired carbon from CH₄ in the sediments of this arctic lake.