Nitrogen removal in a sequencing batch biofilm reactor with liquid circulation: Effect of feed strategy and carbon source in the denitrifying step

Ammonium nitrogen removal from a synthetic wastewater by nitrification and denitrification processes were performed in a sequencing batch biofilm reactor containing immobilized biomass on polyurethane foam with circulation of the liquid-phase. It was analyzed the effect of four external carbon sources (ethanol, acetate, carbon synthetic medium and methanol) acting as electron donors in the denitrifying process. The experiments were conducted with intermittent aeration and operated at 30+/-1 degrees C in 8-h cycles. The synthetic wastewater (100 mgCOD/L and 50 mgNH(4)(+)-N/L) was added batch-wise, while the external carbon sources were added fed-batch-wise during the periods where aeration was suspended. Ammonium nitrogen removal efficiencies obtained were 95.7, 94.3 and 97.5% for ethanol, acetate and carbon synthetic medium, respectively. As to nitrite, nitrate and ammonium nitrogen effluent concentrations, the results obtained were, respectively: 0.1, 5.7 and 1.4 mg/L for ethanol; 0.2, 4.1 and 1.8 mg/L for acetate and 0.2, 6.7 and 0.8 for carbon synthetic medium. On the other hand using methanol, even at low concentrations (50% of the stoichiometric value calculated for complete denitrification), resulted in increasing accumulation of nitrate and ammonium nitrogen in the effluent over time.

Feasibility of a Sequencing Reactor Operated in Batch and Fed-batch Mode Applied to Nitrification and Denitrification Processes

The objective of this work was to study the operational feasibility of nitrification and denitrification processes in a mechanically stirred sequencing batch reactor (SBR) operated in batch and fed-batch mode. The reactor was equipped with a draft-tube to improve mass transfer and contained dispersed (aerobic) and granulated (anaerobic) biomass. The following reactor variables were adjusted: aeration time during the nitrification step; dissolved oxygen concentration, feed time defining batch and fed-batch phases, concentration of external carbon source used as electron donor during the denitrification stage and volumetric ammonium nitrogen load in the influent. The reactor (5 L volume) was maintained at 30 +/- 1 degrees C and treated either 1.0 or 1.5 L wastewater in 8-h cycles. Ammonium nitrogen concentrations assessed were: 50 (condition 1) and 100 mgN-NH(4)(+).L(-1) (condition 2), resulting in 29 and 67 mgN-NH(4)(+).L-1-d(-1), respectively. A synthetic medium and ethanol were used as external carbon sources (ECS). Total nitrogen removal efficiencies were 94.4 and 95.9% when the reactor was operated under conditions 1 and 2, respectively. Low nitrite (0.2 and 0.3 mgN-NO(2)(-).L(-1), respectively) and nitrate (0.01 and 0.3 mgN-NO(3)(-).L(-1), respectively) concentrations were detected in the effluent and ammonium nitrogen removal efficiencies were 97.6% and 99.6% under conditions 1 and 2, respectively.

Influence of cerium (IV) ions on the mechanism of organosilane polymerization and on the improvement of its barrier properties

In this work, the effect of cerium (IV) ammonium nitrate (CAN) addition on the polymerization of bis-[triethoxysilyl]ethane (BTSE) film applied on carbon steel was studied. The electrochemical characterization of the films was carried out in 0.1 mol L(-1) NaCl solution by open-circuit potential measurements, anodic and cathodic polarization curves and electrochemical impedance spectroscopy (EIS). Morphological and chemical characterization were performed by atomic force microscopy (AFM), contact angle measurements, infrared-spectroscopy, nuclear magnetic resonance and thermogravimetric analysis. The results have clearly shown the improvement on the protective properties of the Ce(4+) modified film as a consequence of the formation of a more uniform and densely reticulated silane film. A mechanism is proposed to explain the accelerating role of Ce(4+) ions on the cross-linking of the silane layer. (C) 2008 Elsevier Ltd. All rights reserved.

Carbon and nitrogen cycling in a tropical Brazilian soil cropped with sugarcane and irrigated with wastewater

Carbon (C) and nitrogen (N) dynamics in agro-systems can be altered as a consequence of treated sewage effluent (TSE) irrigation. The present study evaluated the effects of TSE irrigation over 16 months on N concentrations in sugarcane (leaves, stalks and juice), total soil carbon (TC), total soil nitrogen (TN), NO(3)(-)-N in soil and nitrate (NO(3)(-)) and dissolved organic carbon (DOC) in soil solution. The soil was classified as an Oxisol and samplings were carried out during the first productive crop cycle, from February 2005 (before planting) to September 2006 (after sugarcane harvest and 16 months of TSE irrigation). The experiment was arranged in a complete block design with five treatments and four replicates. Irrigated plots received 50% of the recommended mineral N fertilization and 100% (T100), 125% (T125), 150% (T150) and 200% (T200) of crop water demand. No mineral N and irrigation were applied to the control plots. TSE irrigation enhanced sugarcane yield but resulted in total-N inputs(804-1622 kg N ha(-1)) greater than exported N (463-597 kg N ha(-1)). Hence, throughout the irrigation period, high NO(3)(-) concentrations (up to 388 mg L(-1) at T200) and DOC (up to 142 mg L(-1) at T100) were measured in soil solution below the root zone, indicating the potential of groundwater contamination. TSE irrigation did not change soil TC and TN. (C) 2009 Elsevier B.V. All rights reserved.

Leaching of nutrients from a sugarcane crop growing on an Ultisol in Brazil

Leaching is disadvantageous, both for economical and environmental reasons since it may decrease the ecosystem productivity and may also contribute to the contamination of surface and ground water. The objective of this paper was to quantify the loss of nitrogen and sulfur by leaching, at the depth of 0.9 m, in an Ultisol in Sao Paulo State (Brazil) with high permeability, Cultivated with sugarcane during the agricultural cycle of crop plant. The following ions were evaluated: nitrite, nitrate, ammonium, and sulfate. Calcium, magnesium, potassium, and phosphate were also evaluated at the same depth. The sugarcane was planted and fertilized in the furrows with 120 log ha(-1) of N-urea. In order to find out the fate of N-fertilizer, four microplots with (15)N-enriched fertilizer were installed. Input and output of the considered ions at the depth of 0.9 m were quantified from the flux density of water and the concentration of the elements in the soil solution at this soil depth: tensiometers, soil water retention curve and soil solution extractors were used for this quantification. The internal drainage was 205 mm of water, with a total loss of 18 kg ha(-1) of N and 10 kg ha(-1) of S. The percentage of N in the soil solution derived from the fertilizer (%NSSDF) was 1.34, resulting in only 25 g ha(-1) of N fertilizer loss by leaching during all agricultural cycle. Under the experimental conditions of this crop plant, that is, high demand of nutrients and high incorporation of crop residues, the leached N represented 15% of applied N and S leaching were not considerable; the higher amount of leached N was native nitrogen and a minor quantity from N fertilizer; and the leached amount of Ca, Mg, K and P did not exceed the applications performed in the crop by lime and fertilization. (C) 2009 Elsevier B.V. All rights reserved.

Distribution and storage characterization of soil solution for drip irrigation

The increased use of marginal quality water with drip irrigation requires sound fertigation practices that reconcile environmental concerns with viable crop production objectives. We conducted experiments to characterize dynamics and patterns of soil solution within wet bulb formed by drip irrigation. Time-domain reflectometry probes were used to monitor the distribution of potassium nitrate (KNO(3)) and water distribution from drippers discharging at constant flow rates of 2, 4 and 8 L h(-1) in soil-filled containers. Considering results from different profiles, we observed greater solute storage near the dripper decreasing gradually towards the wetting front. About half of the applied KNO(3) solution (48%) was stored in the first layer (0-0.10 m) for all experiments, 29% was stored in the next layer (0.10-0.20 m). Comparing different dripper flow rates, we observed higher solution storage for 4 L h(-1), with 45, 53 and 47% of applied KNO(3) solution accumulating in the first layer (0-0.10 m) for dripper flow rates of 2, 4 and 8 L h(-1), respectively. The results suggest that based on the volume and frequency used in this experiment, it would be advantageous to apply small amounts of solution at more frequent intervals to reduce deep percolation losses of applied water and solutes.

Antioxidant enzyme activity in Acidithiobacillus ferrooxidans LR maintained in contact with chalcopyrite

The total protein content and activity of the enzymes glutathione reductase (GR), superoxide dismutase (SOD) and thioredoxin reductase (TrxR) were evaluated in Acidithiobacillus ferrooxidans LR cells maintained in contact with the metal sulfide chalcopyrite for 1 and 10 days. A significant decrease in total protein content was observed in cells maintained for 10 days in the presence of chalcopyrite, suggesting proteolytic breakdown clue to exposure to the metal sulfide. Following 10 clays of contact with chalcopyrite, increases in GR, SOD and TrxR activities were detected, suggesting the formation of reactive oxygen species. After ten clays, there was a fivefold increase in GR activity, of which, isoenzyme IV represented approximately 82% of the total. An increase in Fe-SOD activity following ten days exposure to chalcopyrite was also determined, as measured on non-denaturing polyacrylamide gels. Also, after 10 days. an approximately 31-fold increase was observed for TrxR activity. The presence of oxidative stress when A. ferrooxidans is in the presence of chalcopyrite could have a negative impact on bioleaching. (C) 2010 Elsevier Ltd. All rights reserved.

Zn uptake, physiological response and stress attenuation in mycorrhizal jack bean growing in soil with increasing Zn concentrations

The influence of arbuscular mycorrhizal fungi (AMF) inoculation on Canavalia ensiformis growth. nutrient and Zn uptake, and on some physiological parameters in response to increasing soil Zn concentrations was studied. Treatments were applied in seven replicates in a 2 x 4 factorial design, consisting of the inoculation or not with the AMF Glomus etunicatum, and the addition of Zn to soil at the concentrations of 0, 100, 300 and 900 mg kg(-1). AMF inoculation enhanced the accumulation of Zn in tissues and promoted biomass yields and root nodulation. Mycorrhizal plants exhibited relative tolerance to Zn up to 300 mg kg(-1) without exhibiting visual symptoms of toxicity, in contrast to non-mycorrhizal plants which exhibited a significant growth reduction at the same soil Zn concentration. The highest concentration of Zn added to soil was highly toxic to the plants. Leaves of plants grown in high Zn concentration exhibited a Zn-induced proline accumulation and also an increase in soluble amino acid contents; however proline contents were lower in mycorrhizal jack beans. Plants in association or not with the AMF exhibited marked differences in the foliar soluble amino acid profile and composition in response to Zn addition to soil. In general, Zn induced oxidative stress which could be verified by increased lipid peroxidation rates and changes in catalase, ascorbate peroxidase, glutathione reductase and superoxide dismutase activities. In summary, G. etunicatum was able to maintain an efficient symbiosis with jack bean plants in moderately contaminated Zn-soils, improving plant performance under those conditions, which is likely to be due to a combination of physiological and nutritional changes caused by the intimate relation between fungus and plant. The enhanced Zn uptake by AMF inoculated jack bean plants might be of interest for phytoremediation purposes. (C) 2009 Elsevier Ltd. All rights reserved.

Biochemical responses of the ethylene-insensitive Never ripe tomato mutant subjected to cadmium and sodium stresses

In order to further address the known interaction between ethylene and components of the oxidative system, we have used the ethylene-insensitive Never ripe (Nr) tomato (Solanum lycopersicum L) mutant, which blocks ethylene responses. The mutant was compared to the control Micro-Tom (MT) cultivar subjected to two stressful situations: 100 mM NaCl and 0.5 mM CdCl(2). Leaf chlorophyll, lipid peroxidation and antioxidant enzyme activities in roots, leaves and fruits, and Na and Cd accumulation in tissues were determined. Although we verified a similar growth pattern and Na and Cd accumulation for MT and Nr, the mutant exhibited reduced leaf chlorophyll degradation following stress. In roots and leaves, the patterns of catalase (CAT), glutathione reductase (GR), ascorbate peroxidase (APX), guaiacol peroxidase (GPOX), superoxide dismutase (SOD) enzyme activity as well as malondialdehyde (MDA) and hydrogen peroxide (H(2)O(2)) production under the stressful conditions tested were very similar between MT and Nr mutant. However, Nr fruits showed increased H(2)O(2) production, reduced and enhanced APX activity in NaCl and CdCl(2), respectively, and enhanced GPOX in NaCl. Moreover, through non-denaturing PAGE, a similar reduction of SOD I band intensity in both, control MT and Nr mutant, treated with NaCl was observed. In leaves and fruits, a similar SOD activity pattern was observed for all periods, genotypes and treatments. Overall the results indicate that the ethylene signaling associated with NR receptor can modulate the biochemical pathways of oxidative stress in a tissue dependent manner, and that this signaling may be different following Na and Cd exposure. (C) 2011 Elsevier B.V. All rights reserved.

The isolation of antioxidant enzymes from mature tomato (cv. Micro-Tom) plants

The activity of catalase (CAT), guaiacol peroxidase (GPOX), ascorbate peroxidase (APX), glutathione reductase (GR), and the isoenzymes of superoxide dismutase (SOD) were determined in the organs of tomato (Lycopersicon esculentum) cultivar Micro-Tom after 104 days of development. The total activities of CAT, GPOX, and GR were higher in the stem than in others tissues, whereas the stem exhibited the lowest APX activity. Activity staining analysis following gel electrophoresis revealed the existence of four SOD isoenzymes in leaves, three in fruits, but only two in the roots and stems. This characterization is essential for an investigation into the effect of abiotic and biotic stresses on the oxidative stress responses by this plant model system.

Effects of the herbicides acetochlor and metolachlor on antioxidant enzymes in soil bacteria

The aim of this study was to investigate the antioxidant responses of three bacteria (SD1. KD and K9) isolated from soil previously treated with the herbicides metolachlor and acetochlor. By 165 rRNA gene sequencing, we determined that SD1 is phylogenetically related to Enterobacter asburiae, while KD and K9 have divergent genomes that more closely resemble that of Enterobacter amnigenus. Decreased levels of lipid peroxidation were observed in SD1 and KD following treatment with 34 mM metolachlor or 62 mM acetochlor, respectively, indicating that both bacteria were able to adapt to an increase in ROS production. In the presence of 34 mM metolachlor or 62 mM acetochlor, all bacterial isolates exhibited increases in total catalase (CAT) activity (81% for SDI, 53% for KD and 59% for K9), whereas total SOD activity (assessed based on the profile and intensity of the bands) was slightly reduced when the bacteria were exposed to high concentrations of the herbicides (340 mM metolachlor or 620 mM acetochlor). This effect was due to a specific reduction in SOD IV (K9 and KD isolates) by 45% and 90%, respectively, and SOD V (SD1 isolate) isoenzymes by 60%. The most striking result was obtained in the SD1 isolate, where two novel isoenzymes of glutathione reductase (GR) that responded specifically to metolachlor were identified. In addition, acetochlor was shown to induce the expression of a new 57 kDa protein band in the K9 and KD isolates. The bacteria isolated from the herbicide-contaminated soil exhibited an efficient antioxidant system response at herbicide concentrations of up to 34 mM metolachlor or 62 mM acetochlor. These data suggest a mechanism for tolerance that may include the control of an imbalance in ROS production versus scavenging. The data suggest that specific isoenzymes of CAT and GR could be involved in this herbicide tolerance mechanism. (C) 2011 Elsevier Ltd. All rights reserved.

Antioxidant response of Nicotiana tabacum cv. Bright Yellow 2 cells to cadmium and nickel stress

Plant cell cultures are a suitable model system for investigation of the physiological mechanisms of tolerance to environmental stress. We have determined the effects of Cd (0.1 and 0.2 mM CdCl(2)) and Ni (0.075 and 0.75 mM NiCl(2)) on Nicotiana tabacum L. cv. Bright Yellow (TBY-2) cell suspension cultures over a 72-h period. Inhibition of growth, loss of cell viability and lipid peroxidation occurred, in general, only when the TBY-2 cells were grown at 0.2 mM CdCl(2) and at 0.75 mM NiCl(2). At 0.1 mM CdCl(2), a significant increase in growth was determined at the end of the experiment. Increases in the activities of all of the four enzymatic antioxidant defence systems tested, were induced by the two concentrations of Cd and Ni, but at different times during the period of metal exposure. Overall, the cellular antioxidant responses to Cd and Ni were similar and were apparently sufficient to avoid oxidative stress at the lower concentrations of Cd and Ni. The activities of glutathione reductase and glutathione S-transferase increased early but transiently, whereas the activities of catalase and guaiacol peroxidase increased in the latter half of the experimental period. Therefore it is likely that the metabolism of reduced glutathione was enhanced during the initial onset of the stress, while catalase and guaiacol-type peroxidase appeared to play a more important role in the antioxidant response once the stress became severe.

Acquired tolerance of tomato (Lycopersicon esculentum cv. Micro-Tom) plants to cadmium-induced stress

The effects of varying concentrations of cadmium (Cd) on the development of Lycopersicon esculentum cv. Micro-Tom (MT) plants were investigated after 40 days (vegetative growth) and 95 days (fruit production), corresponding to 20 days and 75 days of exposure to CdCl(2), respectively. Inhibition of growth was clearly observed in the leaves after 20 days and was greater after 75 days of growth in 1 mM CdCl(2), whereas the fruits exhibited reduced growth following the exposure to a concentration as low as 0.1 mM CdCl(2). Cd was shown to accumulate in the roots after 75 days of growth but was mainly translocated to the upper parts of the plants accumulating to high concentrations in the fruits. Lipid peroxidation was more pronounced in the roots even at 0.05 mM CdCl(2) after 75 days, whereas in leaves, there was a major increase after 20 days of exposure to 1 mM CdCl(2), but the fruit only exhibited a slight significant increase in lipid peroxidation in plants subjected to 1 mM CdCl(2) when compared with the control. Oxidative stress was also investigated by the analysis of four key antioxidant enzymes, which exhibited changes in response to the increasing concentrations of Cd tested. Catalase (EC 1.11.1.6) activity was shown to increase after 75 days of Cd treatment, but the major increases were observed at 0.1 and 0.2 mM CdCl(2), whereas guaiacol peroxidase (EC 1.11.1.7) did not vary significantly from the control in leaves and roots apart from specific changes at 0.5 and 1 mM CdCl(2). The other two enzymes tested, glutathione reductase (EC 1.6.4.2) and superoxide dismutase (SOD, EC 1.15.1.1), did not exhibit any significant changes in activity, apart from a slight decrease in SOD activity at concentrations above 0.2 mM CdCl(2). However, the most striking results were obtained when an extra treatment was used in which a set of plants was subjected to a stepwise increase in CdCl(2) from 0.05 to 1 mM, leading to tolerance of the Cd applied even at the final highest concentration of 1 mM. This apparent adaptation to the toxic effect of Cd was confirmed by biomass values being similar to the control, indicating a tolerance to Cd acquired by the MT plants.

A role for ferritin in the antioxidant system in coffee cell cultures

Iron (Fe) is an essential nutrient for plants, but it can generate oxidative stress at high concentrations. In this study, Coffea arabica L. cell suspension cultures were exposed to excess Fe (60 and 240 mu M) to investigate changes in the gene expression of ferritin and antioxidant enzymes. Iron content accumulated during cell growth, and Western blot analysis showed an increase of ferritin in cells treated with Fe. The expression of two ferritin genes retrieved from the Brazilian coffee EST database was studied. CaFER1, but not CaFER2, transcripts were induced by Fe exposure. Phylogenetic analysis revealed that CaFER1 is not similar to CaFER2 or to any ferritin that has been characterised in detail. The increase in ferritin gene expression was accompanied by an increase in the activity of antioxidant enzymes. Superoxide dismutase, guaiacol peroxidase, catalase, and glutathione reductase activities increased in cells grown in the presence of excess Fe, especially at 60 mu M, while the activity of glutathione S-transferase decreased. These data suggest that Fe induces oxidative stress in coffee cell suspension cultures and that ferritin participates in the antioxidant system to protect cells against oxidative damage. Thus, cellular Fe concentrations must be finely regulated to avoid cellular damage most likely caused by increased oxidative stress induced by Fe. However, transcriptional analyses indicate that ferritin genes are differentially controlled, as only CaFER1 expression was responsive to Fe treatment.

Nitrogen and Sulfur Fertilization and Dynamics in a Brazilian Entisol under Pasture

Nutrient dynamics in tropical soils sustaining forage grasses are still poorly understood. We conducted a study to evaluate the effect of combined N and S fertilizer rates on the growth of `Marandu` palisade grass [Brachiaria brizantha (Hochst. ex A. Rich.) Stapf], uptake of these elements from the soil by plants, soil organic matter concentration, soil pH, and the mineral and organic fractions of N and S in an Entisol. Combinations of five N rates (0, 100, 200, 300, and 400 g N m(-3)) with five S rates (0, 10, 20, 30, and 40 g S m(-3)) were evaluated in a partial 5 x 5 factorial in a pot experiment, with and without plants. Nitrogen and S were supplied as NH(4)NO(3) and CaSO(4)center dot 2H(2)O, respectively. The N addition in excess did not enhance the palisade grass production due to low plant-available Sin the soil. The supply of low rates of S with N greatly improved the overall N uptake efficiency by the forage plant. The contents of total N, NO(3)(-)-N, and NH(4)(+)-N in the soil varied with N rate and with N uptake by the plants. The association of palisade grass with S fertilization increased the ester-bonded S fraction in the soil. The results suggest that soil residual S could be a potential source of S for plants. Proper N and S fertilizer rates promoted increased grass production due to increased uptake of these nutrients and the dynamics of the organic N and S fractions and mineral fractions in this tropical soil.