Copper enhances the activity and salt resistance of mixed methane-oxidizing communities

Effluents of anaerobic digesters are an underestimated source of greenhouse gases, as they are often saturated with methane. A post-treatment with methane-oxidizing bacterial consortia could mitigate diffuse emissions at such sites. Semi-continuously fed stirred reactors were used as model systems to characterize the influence of the key parameters on the activity of these mixed methanotrophic communities. The addition of 140 mg L⁻¹ NH₄⁺–N had no significant influence on the activity nor did a temperature increase from 28°C to 35°C. On the other hand, addition of 0.64 mg L⁻¹ of copper(II) increased the methane removal rate by a factor of 1.5 to 1.7 since the activity of particulate methane monooxygenase was enhanced. The influence of different concentrations of NaCl was also tested, as effluents of anaerobic digesters often contain salt levels up to 10 g NaCl L⁻¹. At a concentration of 11 g NaCl L⁻¹, almost no methane-oxidizing activity was observed in the reactors without copper addition. Yet, reactors with copper addition exhibited a sustained activity in the presence of NaCl. A colorimetric test based on naphthalene oxidation showed that soluble methane monooxygenase was inhibited by copper, suggesting that the particulate methane monooxygenase was the active enzyme and thus more salt resistant. The results obtained demonstrate that the treatment of methane-saturated effluents, even those with increased ammonium (up to 140 mg L⁻¹ NH₄⁺–N) and salt levels, can be mitigated by implementation of methane-oxidizing microbial consortia.     

Change in ammonia-oxidizing microorganisms in enriched nitrifying activated sludge

In this study, sludge was taken from a municipal wastewater treatment plant that contained a nearly equal number of archaeal amoA genes (5.70 × 10⁶ ± 3.30 × 10⁵ copies mg sludge⁻¹) to bacterial amoA genes (8.60 × 10⁶ ± 7.64 × 10⁵ copies mg sludge⁻¹) and enriched in three continuous-flow reactors receiving an inorganic medium containing different ammonium concentrations: 2, 10, and 30 mM NH₄⁺–N (28, 140, and 420 mg N l⁻¹). The abundance and communities of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in enriched nitrifying activated sludge (NAS) were monitored at days 60 and 360 of the operation. Early on, between day 0 and day 60 of reactor operation, comparative abundance of AOA amoA genes to AOB amoA genes varied among the reactors depending on the ammonium levels found in the reactors. As compared to the seed sludge, the number of AOA amoA genes was unchanged in the reactor with lower ammonium level (0.06 ± 0.04 mgN l⁻¹), while in the reactors with higher ammonium levels (0.51 ± 0.33 and 0.25 ± 0.10 mgN l⁻¹), the numbers of AOA amoA genes were deteriorated. By day 360, AOA disappeared from the ammonia-oxidizing consortiums in all reactors. The majority of the AOA sequences from all NASs at each sampling period fell into a single AOA cluster, however, suggesting that the ammonium did not affect the AOA communities under this operational condition. This result is contradictory to the case of AOB, where the communities varied significantly among the NASs. AOB with a high affinity for ammonia were present in the reactors with lower ammonium levels, whereas AOB with a low affinity to ammonia existed in the reactors with higher ammonium levels.     

A preliminary study of the bioremediation potential of Codium fragile applied to seaweed integrated multi-trophic aquaculture (IMTA) during the summer

In integrated multi-trophic aquaculture (IMTA), seaweeds have the capacity to reduce the environmental impact of nitrogen-rich effluents in coastal ecosystems. To establish such bioremediation systems, selection of suitable seaweed species is important. The distribution and productivity of seaweeds vary seasonally based on water temperature and photoperiod. In Korea, candidate genera such as Pophyra, Laminaria, and Undaria grow from autumn to spring. In contrast, Codium grows well at relatively high water temperatures in summer. Thus, aquaculture systems potentially could capitalize on Codium’s capacity for rapid growth in the warm temperatures of late summer and early fall. In this study, we investigated ammonium uptake and removal efficiency by Codium fragile. In laboratory experiments, we grew C. fragile under various water temperatures (10, 15, 20, and 25°C), irradiances (dark, 10, and 100 μmol photons m⁻² s⁻¹), and initial ammonium concentrations (150 and 300 μM); in all cases, C. fragile exhausted the ammonium supply for 6 h. At 150 μM of , ammonium removal efficiency was greatest (99.5 ± 2.6%) when C. fragile was incubated at 20°C under 100 μmol photons m⁻² s⁻¹. At 300 μM of , removal efficiency was greatest (86.3 ± 2.1%) at 25°C under 100 μmol photons m⁻² s⁻¹. Ammonium removal efficiency was significantly greater at 20 and 25°C under irradiance of 100 μmol photons m⁻² s⁻¹ than under other conditions tested.     

Suppression of nitrate formation within an exotic conifer plantation

Summary Nitrate-N losses to stream waters and soil inorganic N pools, nitrifying potentials and NO₃-N production rates were measured in 2 adjacent watersheds, one used as pasture and the other planted in exotic conifer forest (Pinus radiata D. Don). Estimated NO₃-N loss to stream waters draining the pine and pasture watersheds were 0.6kg ha⁻¹ y⁻¹ and 7.6 kg ha⁻¹ y⁻¹ respectively. Ammonium-N pool sizes were not significantly different between soils in the two watersheds but NO₃−N pools and nitrifying potentials were always lower in the pine watershed soil samples. Laboratory incubation experiments indicated that suppression of NO₃−N formation in pine watershed soils required the presence of live tree roots and was not due to the direct action of allelopathic chemicals on nitrifiers.     

A Novel Approach for Bioremediation of a Coastal Marine Wastewater Effluent Based on Artificial Microbial Mats

A novel alternative for wastewater effluent bioremediation was developed using constructed microbial mats on low-density polyester. This biotechnology showed high removal efficiencies for nitrogen and phosphorous in a short retention time (48 h): 94% for orthophosphate (7.78 g m³ d⁻¹), 79% for ammonium (11.30 g m⁻³ d⁻¹), 78% for nitrite (7.46 g m⁻³ d⁻¹), and 83% for nitrate (8.55 g m⁻³ d⁻¹). The microbial mats were dominated by Cyanobacteria genera such as Chroococcus sp., Lyngbya sp., and bacteria of the subclass Proteobacteria representative of the Eubacteria Domain. Nitzschia sp. was the dominant Eukaryote Domain. Various N and P substrates in the wastewater permit the growth of self-forming and self-sustaining bacterial, microalgal, and cyanobacterial communities on a polyester support. The result is the continuous, self-sufficient growth of microbial mats. This is an innovative, economical, and environmentally safe alternative for the treatment of wastewater effluents in coastal marine environments.     

Activity of Nitrifying Biofilms Constructed on Low-Density Polyester Enhances Bioremediation of a Coastal Wastewater Effluent

Summary Nitrifying biofilms were constructed on low density polyester Dacron for the bioremediation of nitrogen from wastewater effluent of a municipal treatment plant. Dacron disks were inoculated with wastewater sludge enriched for 15 days for either ammonia- or nitrite-oxidizing bacteria (AOB or NOB, respectively) and packed into glass bioreactors. Wastewater effluent containing high levels of ammonia, nitrite, and phosphate was collected and fed to inoculated and uninoculated bioreactors. Both inoculated bioreactors showed stable nitrification efficiencies, removing 96 and 76% of the ammonia and 12 and 35% of the nitrite for AOB- and NOB-inoculated bioreactors, respectively. Efficiencies of phosphate removal were similar in both inoculated and uninoculated bioreactors, indicating that nitrifiers were not required for this process. AOB-inoculated bioreactors accumulated nitrite mid-way through the experiment and had low rates of conversion to nitrate, suggesting slow nitrite oxidizer growth. DGGE and sequence analysis of AOB 16S rRNA genes showed enrichment of Nitrosomonas spp. in both inoculated bioreactors, and a dominance of Nitrosospira spp. in non-inoculated bioreactors. This study describes an inexpensive and efficient technology for removing ammonia and nitrite from wastewater effluents of municipal treatment plants before its release to the environment.     

Characterization of microbial community and kinetics for spent sulfidic caustic applied autotrophic denitrification

Spent sulfidic caustic was applied to sulfur utilizing autotrophic denitrification as the simultaneous source of electron donor and alkalinity. The two experiment set-up of upflow anoxic hybrid growth reactor (UAHGR) and upflow anoxic suspended growth reactor (UASGR) was adopted and nitrate removals were similar in both reactors. Approximately 90% of the initial nitrate was denitrified at nitrate loading rate of 0.15∼0.40 kgNO₃ ⁻/m³·d. The experimental stoichiometric ratio of sulfate production to nitrate removal was ranged from 1.5 to 2.1 mgSO₄ ²⁻/mgNO₃ ⁻. During the operation period, denaturing gradient gel electrophoresis (DGGE) analysis of polymerase chain reaction (PCR)-amplified 16S rDNA fragments for the sludge sample of both reactors showed the change of microbial communities. Thiobacillus denitrificans-like microorganism occupied 28.5% (18 clones) of the 63 clones by cloning the PCR products from the sludge sample of UAHGR. Acidovorax avenae, which can reduce nitrate to nitrogen gas while oxidizing phenol (heterotrophic denitrifier), was also found in 7 clones (11.1%). Although an organic carbon source was not added to the medium, a microorganism (Kaistella koreensis) capable of oxidizing organic compounds was found in 7 clones (11.1%). Therefore, the microbial community of spent sulfidic caustic applied autotrophic denitrification process well corresponds to the substrate components of spent sulfidic caustic. Through the batch cultivation of microorganisms in UAHGR, the microbial kinetic coefficients of spent sulfidic caustic applied autotrophic denitrification were estimated to be μ _max = 0.097 h⁻¹, k _d = 0.0021 h⁻¹, K _s = 200 mgNO₃ ⁻/L, and Y = 0.31 mgMLVSS/mgNO₃ ⁻.     

Measurement of potential activity of fixed nitrifying bacteria in biological filters used in drinking water production

Nitrification during biological filtration is being used more and more in drinking water production to remove ammonia, which can be the source of several water quality problems during distribution. In this process, ammonia is converted into nitrite and then into nitrate by fixed autotrophic nitrifying bacteria. The purpose of this work was to develop a technique to estimate fixed nitrifying biomass (sum of ammonia- and nitrite-oxidizing populations). The quantification of autotrophic nitrifying biomass was determined by potential nitrifying activity measurement. The production of oxidized forms of inorganic nitrogen (nitrates and nitrites) was measured after an incubation of 2 cm³ of colonized solid support in the presence of a 5-ml nitrifier medium containing 10 mg N-NH₄ L⁻¹ for 30 min at 32°C. The production rate of oxidized nitrogen in optimal conditions was measured and converted into nitrifying biomass by using the maximum specific oxidizing activity. This technique was shown to be appropriate for conditions encountered in the biological filters used in drinking water production and sufficiently simple to be used for routine measurements. Journal of Industrial Microbiology & Biotechnology (2000) 24, 161–166. Received 28 July 1999/ Accepted in revised form 11 November 1999     

Binding of nitrite-N on polyphenols during nitrification

Summary The conversion of substantial amounts of ammonia nitrogen into organic nitrogen as a direct result of nitrification at neutral pH-values, was monitored in soil suspensions amended with ammonium nitrogen. The influence of the chemolithotrophic nitrifying bacteria was verified by applying nitrapyrin as a selective inhibitor in control experiments. In addition, the role of phenolic compounds was examined by adding α naphthol. The factors influencing the nitrification processi.e. pH, NH ₄ ⁺ −N, NO ₂ ⁻ −N, NO ₃ ⁻ −N were measured during a 60 days incubation period. Nitrification started to be active after 5 and 10 days in the normal and the naphthol spiked soil suspensions respectively; it was inhibited in the nitrapyrin controls. Parallel with nitrification, formation of organic nitrogen was observed. The humic matter fractions were extracted and analyzed by I.R. spectroscopy which revealed the valence vibration ranges of nitro and nitroso groups fixed in different positions on aromatic compounds, both for normal and naphthol spiked samples. High resolution gas chromatography combined with mass spectroscopic analysis indicated the formation of nitrosonaphtholes. In addition a novel organic nitro compound was identifiedi.e. an azido nitro benzene. No nitrogen was fixed in the samples treated with nitrification inhibitor. A mechanism for the fixation of nitrite nitrogen during nitrification is proposed.     

Modeling nitrogen cycling in a coastal fresh water sediment

Increased nitrogen (N) loading to coastal marine and freshwater systems is occurring worldwide as a result of human activities. Diagenetic processes in sediments can change the N availability in these systems, by supporting removal through denitrification and burial of organic N (N_org) or by enhancing N recycling. In this study, we use a reactive transport model (RTM) to examine N transformations in a coastal fresh water sediment and quantify N removal rates. We also assess the response of the sediment N cycle to environmental changes that may result from increased salinity which is planned to occur at the site as a result of an estuarine restoration project. Field results show that much of the N_org deposited on the sediment is currently remineralized to ammonium. A rapid removal of nitrate is observed in the sediment pore water, with the resulting nitrate reduction rate estimated to be 130 μmol N cm⁻² yr⁻¹. A model sensitivity study was conducted altering the distribution of nitrate reduction between dissimilatory nitrate reduction to ammonium (DNRA) and denitrification. These results show a 40% decline in sediment N removal as NO ₃ ⁻ reduction shifts from denitrification to DNRA. This decreased N removal leads to a shift in sediment-water exchange flux of dissolved inorganic nitrogen (DIN) from near zero with denitrification to 133 μmol N cm⁻² yr⁻¹ if DNRA is the dominant pathway. The response to salinization includes a short-term release of adsorbed ammonium. Additional changes expected to result from the estuarine restoration include: lower NO ₃ ⁻ concentrations and greater SO ₄ ²⁻ concentrations in the bottom water, decreased nitrification rates, and increased sediment mixing. The effect of these changes on net DIN flux and N removal vary based on the distribution of DNRA versus denitrification, illustrating the need for a better understanding of factors controlling this competition.     

Nitrification, denitrification, and dechlorination in bleached kraft pulp mill wastewater

This study deals with combining the biologi cal removal of organic halogens with the removal of nitrogen from bleached kraft pulp mill wastewater in fluidized-bed reactors under nitrifying and denitrifying conditions. Untreated and biotreated bleached kraft pulp mill wastewaters had no detrimental effect on nitrification or denitrification. The nitrifying biofilm reactor, pregrown on synthetic inorganic feed with ammonia, removed without a lag phase adsorbable organic halogens [7.2 mg Cl (g biomass volatile solids)⁻¹day⁻¹] from bleached kraft pulp mill wastewater and selected chlorophenols from synthetic wastewater. Electron microscopical examination of the biofilm showed that bacteria, morphologically similar to the nitrifying species Nitrosomonas or Nitrobacter, and Nitrosospira were dominant. The denitrifying fluidized-bed reactor, pregrown on nitrate and methanol, denitrified without a lag phase bleached kraft pulp mill wastewater. Under denitrifying conditions, 35% of the total organic carbon content of untreated bleached kraft pulp mill waste water was removed. The reducing power delivered by untreated bleached kraft pulp mill wastewater for denitrification was 2 mmol electrons/mmol carbon mineralized. Dechlorination under denitrifying conditions was negligible. Received: 21 November 1996 / Received revision: 27 January 1997 / Accepted: 1 February 1997     

Screening and characterization of an aerobic nitrifying-denitrifying bacterium from activated sludge

Taking advantage of the good biocompatibility and high efficiency of nitrogen removal with microbes, nitrifying and denitrifying bacteria, are becoming increasingly more widely used for wastewater treatment and prevention of eutrophication. In this research, an aerobic nitrifying-denitrifying bacterium was successfully screened from activated sludge and identified as Pseudomonas sp. (CCTCC No M2010209) by the 16S rDNA sequence. The activity verification confirmed its nitrifying-denitrifying capability of removing ammonium, nitrate and nitrite nitrogen. The types of carbon sources and carbon-nitrogen ratio greatly influenced the removal efficiency of NH₄ ⁺-N and NO₃ ⁻-N. When the initial concentrations of NH₄ ⁺-N and NO₃ ⁻-N in synthetic wastewater were less than 70 and 50 mg/L, the nitrogen removal rates reached 94 and 90% in 9 h, respectively. Preliminary comparisons of nitrogen removal capacity between this isolate and other commercial preparations in the treatment of synthetic wastewater revealed its promising potential to be used in the actual wastewater treatment.     

Effect of temperature and irradiance on the uptake of ammonium and nitrate by <Emphasis Type="Italic">Laurencia brongniartii</Emphasis> (Rhodophyta,Ceramiales)

The effects of temperature (20, 24 and 28 °C) and irradiance (15 and 40 μmol photon m⁻² s⁻¹) on the nitrate and ammonium uptake rates of the subtropical red alga, Laurencia brongniartii, were investigated to prepare for tank cultivation. Nitrate uptake followed saturation kinetics and was faster at higher irradiances and temperatures. In contrast, ammonium uptake was linear over the experimental range and was not affected by an increase in temperature. A parameter, β, was calculated to compare substrate uptake rates of nitrate along the linear portion of the uptake curve with that of ammonium. For nitrate, β was lower at low irradiance and higher at high irradiance (β = 0.007 ± 0.003 and 0.030 ± 0.002 [μmol N L⁻¹ (μmol N g_ww⁻¹ d⁻)⁻¹], respectively). However, β was 0.023 ± 0.002 and 0.034 ± 0.002 [μmol N L⁻¹ (μmol N g_ww⁻¹ d⁻¹)⁻¹] for ammonium, suggesting a preference for ammonium over nitrate.     

Propene removal from synthetic waste gas using a hollow-fibre membrane bioreactor

 Hollow-fibre modules containing microporous membrane material were evaluated as bioreactors for waste gas treatment. The reactors were inoculated with the propene-utilizing strain Xanthobacter Py2, which formed a biofilm on the inner side of the fibres. The removal of the poorly soluble volatile propene from synthetic waste gas was monitored for up to 170 days. The maximum removal rates were 70–110 g propene per m³ reactor per hour. A gas residence time of 80 s was required to remove 95% of an initial propene concentration of 0.84 g/m³. The presence of ammonium in the liquid medium resulted in the development of an additional population of nitrifying organisms. Therefore, nitrate was used as the source of nitrogen in later experiments. During long-term operation, the propene removal rates gradually decreased. At low liquid velocities (1–5 cm/s) clogging of individual fibres with excess biomass was observed. Elevation of the liquid velocity in the fibres to 90 cm/s resulted in the formation of a dense biofilm and prevented clogging of the fibres. However, also at this high liquid velocity a gradual decrease in propene removal rate was observed. These results suggest that aging of biofilms is a very important factor in long-term operation of hollow-fibre bioreactors. Received: 24 November 1995 / Received revision: 14 February 1996 / Accepted: 20 February 1996     

Low Nitrification Rates in Acid Scots Pine Forest Soils Are Due to pH-Related Factors

In a previous study, ammonia-oxidizing bacteria (AOB)-like sequences were detected in the fragmentation layer of acid Scots pine (Pinus sylvestris L.) forest soils (pH 2.9–3.4) with high nitrification rates (>11.0 μg g⁻¹ dry soil week⁻¹), but were not detected in soils with low nitrification rates (<0.5 μg g⁻¹ dry soil week⁻¹). In the present study, we investigated whether this low nitrification rate has a biotic cause (complete absence of AOB) or an abiotic cause (unfavorable environmental conditions). Therefore, two soils strongly differing in net nitrification were compared: one soil with a low nitrification rate (location Schoorl) and another soil with a high nitrification rate (location Wekerom) were subjected to liming and/or ammonium amendment treatments. Nitrification was assessed by analysis of dynamics in NH₄ ⁺-N and NO₃ ⁻-N concentrations, whereas the presence and composition of AOB communities were assessed by polymerase chain reaction–denaturing gradient gel electrophoresis and sequencing of the ammonia monooxygenase (amoA) gene. Liming, rather than ammonium amendment, stimulated the growth of AOB and their nitrifying activity in Schoorl soil. The retrieved amoA sequences from limed (without and with N amendment) Schoorl and Wekerom soils exclusively belong to Nitrosospira cluster 2. Our study suggests that low nitrification rates in acidic Scots pine forest soils are due to pH-related factors. Nitrosospira cluster 2 detected in these soils is presumably a urease-positive cluster type of AOB.     

Inhibition of Nitrification Alters Carbon Turnover in the Patagonian Steppe

Human activities are altering biodiversity and the nitrogen (N) cycle, affecting terrestrial carbon (C) cycling globally. Only a few specialized bacteria carry out nitrification—the transformation of ammonium (NH ₄ ⁺ ) to nitrate (NO ₃ ⁻ ), in terrestrial ecosystems, which determines the form and mobility of inorganic N in soils. However, the control of nitrification on C cycling in natural ecosystems is poorly understood. In an ecosystem experiment in the Patagonian steppe, we inhibited autotrophic nitrification and measured its effects on C and N cycling. Decreased net nitrification increased total mineral N and NH ₄ ⁺ and reduced NO ₃ ⁻ in the soil. Plant cover (P < 0.05) and decomposition (P < 0.0001) decreased with inhibition of nitrification, in spite of increases in NH ₄ ⁺ availability. There were significant changes in the natural abundance of δ¹⁵N in the dominant vegetation when nitrification was inhibited suggesting that a switch occurred in the form of N (from NO ₃ ⁻ to NH ₄ ⁺ ) taken up by plants. Results from a controlled-condition experiment supported the field results by showing that the dominant plant species of the Patagonian steppe have a marked preference for nitrate. Our results indicate that nitrifying bacteria exert a major control on ecosystem functioning, and that the inhibition of nitrification results in significant alteration of the C cycle. The interactions between the C and N cycles suggest that rates of C cycling are affected not just by the amount of available N, but also by the relative availability for plant uptake of NH ₄ ⁺ and NO ₃ ⁻ .     

NO<Subscript>3</Subscript><Superscript>−</Superscript>-Driven SO<Subscript>4</Subscript><Superscript>2−</Superscript> Production in Freshwater Ecosystems: Implications for N and S Cycling

Massive anthropogenic acceleration of the global nitrogen (N) cycle has stimulated interest in understanding the fate of excess N loading to aquatic ecosystems. Nitrate (NO₃ ⁻) is traditionally thought to be removed mainly by microbial respiratory denitrification coupled to carbon (C) oxidation, or through biomass assimilation. Alternatively, chemolithoautotrophic bacterial metabolism may remove NO₃ ⁻ by coupling its reduction with the oxidation of sulfide to sulfate (SO₄ ²⁻). The NO₃ ⁻ may be reduced to N₂ or to NH₄ ⁺, a form of dissimilatory nitrate reduction to ammonium (DNRA). The objectives of this study were to investigate the importance of S oxidation as a NO₃ ⁻ removal process across diverse freshwater streams, lakes, and wetlands in southwestern Michigan (USA). Simultaneous NO₃ ⁻ removal and SO₄ ²⁻ production were observed in situ using modified “push-pull” methods in nine streams, nine wetlands, and three lakes. The measured SO₄ ²⁻ production can account for a significant fraction (25–40%) of the overall NO₃ ⁻ removal. Addition of ¹⁵NO₃ ⁻ and measurement of ¹⁵NH₄ ⁺ production using the push–pull method revealed that DNRA was a potentially important process of NO₃ ⁻ removal, particularly in wetland sediments. Enrichment cultures suggest that Thiomicrospira denitrificans may be one of the organisms responsible for this metabolism. These results indicate that NO₃ ⁻-driven SO₄ ²⁻ production could be widespread and biogeochemically important in freshwater sediments. Removal of NO₃ ⁻ by DNRA may not ameliorate problems such as eutrophication because the N remains bio-available. Additionally, if sulfur (S) pollution enhances NO₃ ⁻ removal in freshwaters, then controls on N processing in landscapes subject to S and N pollution are more complex than previously appreciated. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Ammonium nutrition increases photosynthesis rate under water stress at early development stage of rice (Oryza sativa L.)

An hydroponic experiment with a simulated water stress induced by PEG (6000) was conducted in a greenhouse to study the effects of nitrate (NO₃ ⁻), ammonium (NH₄ ⁺) and the mixture of NO₃ ⁻ and NH₄ ⁺, on water stress tolerance of rice seedlings. Rice (Shanyou 63) was grown under non- or simulated water stress condition (10% (w/v) PEG, MW6000) with the 3 different N forms during 4 weeks. Under non-stressed condition no difference was observed among the N treatments. Under simulated water stress, seedlings grown on N-NO₃ ⁻ were stunted. Addition of PEG did not affect rice seedling growth in the treatment of only NH₄ ⁺ supply but slightly inhibited the rice seedling growth in the treatment of mixed supply of NO₃ ⁻ and NH₄ ⁺. Simulated water stress, when only N-NH₄ ⁺ was present, did not affect leaf area and photosynthesis rate, however, both parameters decreased significantly in the NO₃ ⁻ containing solutions. Under water stress, Rubisco content in newly expanded leaves significantly increased in the sole NH₄ ⁺ supplied plants as compared to that in plants of the other two N treatments. Under water stress, the ratio of carboxylation efficiency to Rubisco content was, respectively, decreased by 13 and 23% in NH₄ ⁺ and NO₃ ⁻ treatments, respectively. It is concluded that, water stress influenced the Rubisco activity than stomatal limitation, and this effects could be regulated by N forms. Responsible Editor: Herbert Johannes Kronzucker. Shiwei Guo and Gui Chen contributed equally to this paper.     

Seasonal accumulation and partitioning of nitrogen by lentil

Although wheat (Triticum aestivum L.) is the dominant crop of the semi-arid plains of Canada and the western United States, lentil (Lens culinaris Medik.) has become an important alternative crop. Sources and seasonal accumulation of N must be understood in order to identify parameters that can lead to increased N₂-fixing activity and yield. Inoculated lentil was grown in a sandy-loam soil at an irrigated site in Saskatchewan, Canada. Wheat was used as the reference crop to estimate N₂ fixation by the A-value approach. Lentil and wheat received 10 and 100 kg N ha⁻¹ of ammonium nitrate, respectively. Crops were harvested six times during the growing season and plant components analyzed. During the first 71 days after planting the wheat had a higher daily dry matter and N accumulation compared to lentil. However, during the latter part of the growing season, daily dry matter and N accumulation were greater for lentil. The maximum total N accumulation for lentil at maturity was 149 kg ha⁻¹. In contrast, wheat had a maximum N accumulation of 98 kg ha⁻¹ in the Feekes 11.1 stage, or 86 days after planting. The maximum daily rates of N accumulation were 3.82 kg N ha⁻¹ day⁻¹ for lentil and 2.21 kg N ha⁻¹ day⁻¹ for wheat. The percentage of N derived from N₂ fixation (% Ndfa) ranged from 0 at the first harvest to 92 % at final harvest. Generative plant components had higher values for % Ndfa than the vegetative components which indicates that N in the reproductive plant parts was derived largely from current N₂ fixation and lentil continued to fix N until the end of the pod fill stage. At final harvest, lentil had derived 129 kg N ha⁻¹ from N₂ fixation with maximum N₂-fixing activity (4.4 kg N ha⁻¹ day⁻¹) occurring during the early stages of pod fill. Higher maximum rates of N₂-fixing activity than net N accumulation (3.82 kg N ha⁻¹ day⁻¹) may have been caused by N losses like volatilization. In addition, lentil provided a net N contribution to the soil of 59 kg ha⁻¹ following the removal of the grain.     

Chromate reduction by PVA-alginate immobilized <Emphasis Type="Italic">Streptomyces griseus</Emphasis> in a bioreactor

Microbial reduction of toxic Cr⁶⁺ to the less toxic Cr³⁺ is potentially a useful bioremediation process. Among the matrices tested for whole cell immobilization of an efficient chromate-reducing Streptomyces griseus strain, PVA-alginate was the most effective and was used for reduction of Cr(VI) in a bioreactor. Cr⁶⁺ reduction efficiency decreased as Cr⁶⁺ was increased from 2 to 12 mg l⁻¹ but increased with an increase in biomass concentration. However, increasing the flow rate from 2 to 8 ml h⁻¹ did not significantly affect Cr⁶⁺ reduction. The reduction was faster in simulated effluent than in synthetic medium and complete removal of 8 mg Cr⁶⁺ l⁻¹ from effluent and synthetic medium occurred in 2 and 12 h, respectively. Our results indicate that immobilized S. griseus cells could be applied for the large-scale bioremediation of chromate-containing effluents and wastewaters.