Carbon monoxide as an inhibitor of N2ase activity(C2H2) in control measurements of endogenous formation of ethylene by forest soils

Two problems with the recently suggested method to measure endogenous formation of C₂H₄ in an atmosphere enriched with C₂H₂ and CO in studies of N₂ase activity (C₂H₂) in forest soils were analysed, namely the effect of consumption of CO during incubation and the effect of water-saturated conditions.After an initial addition of 100 ml C₂H₂ and 20 ml CO 1^?1 to soil incubation vessels, CO was gradually consumed and followed by a recovery of N₂ase activity when the concentration of CO was lower than about 10 ml 1^?1. The shortest period within which this concentration was achieved was 1 day when incubating fresh soil cores at 15°C, and it was concluded that longer incubations should be avoided.The inhibition of N₂ase activity by CO was strongly suppressed when all soil pores were filled with water. Dissolved inorganic N (0.1% of dry mass soil) was much more efficient in inhibiting N₂ase activity under such conditions.     

Acetylene reduction in soil and litter from pine and eucalypt forests in south-eastern Australia

Rates of C₂H₂-reduction in surface soil and litter from pine and eucalypt forests were measured for 1 yr. Rates of reduction increased significantly with moisture content, and mean rates (nmol kg^?1 h^?1) decreased in the order pine litter (339), eucalypt litter (220), eucalypt soil (54), pine soil (7). Asymbiotic N₂-fixation in litter and surface soil was estimated to be 108 mg m^?2 yr^?1 in eucalypt forest and 64 mg m^?2 yr^?1 in pine forest. About 80% of total fixation in eucalypt was in the soil, while 80% of the total in pine was in the litter. N₂ase was active in rotting wood but not in fresh foliage.     

Nitrogen transformation rates and N<Subscript>2</Subscript>O producing pathways in two pasture soils

Purpose

Better understanding of N transformations and the regulation of N₂O-related N transformation processes in pasture soil contributes significantly to N fertilizer management and development of targeted mitigation strategies.

Materials and methods

¹⁵N tracer technique combined with acetylene (C₂H₂) method was used to measure gross N transformation rates and to distinguish pathways of N₂O production in two Australian pasture soils. The soils were collected from Glenormiston (GN) and Terang (TR), Victoria, Australia, and incubated at a soil moisture content of 60% water-filled pore space (WFPS) and at temperature of 20 °C.

Results and discussion

Two tested pasture soils were characterized by high mineralization and immobilization turnover. The average gross N nitrification rate (n_tot) was 7.28 mg N kg^?1 day^?1 in TR soil () and 5.79 mg N kg^?1 day^?1 in GN soil. Heterotrophic nitrification rates (n_h), which accounting for 50.8 and 41.9% of n_tot, and 23.4 and 30.1% of N₂O emissions in GN and TR soils, respectively, played a role similar with autotrophic nitrification in total nitrification and N₂O emission. Denitrification rates in two pasture soils were as low as 0.003–0.004 mg N kg^?1 day^?1 under selected conditions but contributed more than 30% of N₂O emissions.

Conclusions

Results demonstrated that two tested pasture soils were characterized by fast N transformation rates of mineralization, immobilization, and nitrification. Heterotrophic nitrification could be an important NO₃^?–N production transformation process in studied pasture soils. Except for autotrophic nitrification, roles of heterotrophic nitrification and denitrification in N₂O emission in two pasture soils should be considered when developing mitigation strategies.

     

Nitrogenase activity associated with pasture grasses in Northern New South Wales

Nitrogenase activity associated with roots of grasses was initially examined at 67 sites in New South Wales using an enriched malate medium. Twenty six of the 39 grass species examined produced at least 10 nmol C₂H₄cm^?1 root h^?1—a level accepted as positive presumptive evidence of N₂-fixation: 40 of the 288 samples exceeded 100nmol C₂H₄h^?1. The seasonal N₂ase activity of up to 4 grass species collected in soil cores at 6 sites was measured over 16 months.Activity at field moisture levels, but incubated at 30°C was greatest for cores collected in summer months. Activity was increased in 33.5% of samples by raising soil moisture to field capacity. No single species of grass consistently supported higher activity than any other.Nitrogenase activity was compared in cores of Kikuyu grass (Pennisetum clandestinum) watered to in excess of field capacity and allowed to drain for between 7 and 28 days before re-watering. Activity declined rapidly in the first 7 days and although recovery was also rapid, integration of N₂ase activity over time showed a loss of 20 and 61% for 7 and 28 days drainage respectively. N₂ase activity was greatest at 30°C.Maximum N₂ase activity in field samples was only 246 nmol C₂H₄ core^?1 h^?1 indicating that fixation of N₂ would not be of agronomic significance.     

Influence of sea salts on the soil metabolism

Abstract

Microbial metabolism in reduction process of waterlogged paddy soils has been studied by Takai, Koyama, and Kamura (1, 2, 3, 4, 5, 6), Koyama (7, 8, 9, 10, 11, 12), and others. The results indicated that microbial metabolism in waterlogged soils takes place according to the following steps: (1) In the early stage of the incubation period, dissolved O₂, is consumed and the redox potential drops rapidly. (2) NO₂? and NO₂? are reduced to N₂. (3) Mn⁴⁺ is reduced to Mn²⁺. (4) Fe³⁺ is reduced to Fe²⁺. (5) SO₄ ²? is reduced to S^2?. (6) H₂ and CH₄ are produced. Takai and Chiang (13) reported that NH₄₊ and PO₄ ³⁺ in waterlogged paddy soils increase with the incubation period. Chiang and Takai (14) indicated that carbohydrates in the soil solutions almost remain constant throughout the incubation period, however, organic acids change similarly to those reported previously (5, 6).     

Persistence of glyphosate in a sandy loam

Glyphosale was added to samples of a sandy loam at rates of 0, 2, 5 and 10μg g^?1soil. After 120 days, soil was transferred to pots which were planted with subterranean clover. Plants were inoculated with Rhizohium trifolii and N₂ fixation (C₂H₂-reduction) was recorded after 9, 13, 15 and 19 weeks of growth. Nodule numbers and root weights were determined after the final C₂H₂-reduction assays had been performed. Decreased C₂H₂-reduction, nodule numbers and root weights associated with plants growing in glyphosate-treated soil indicated that this herbicide was not inactivated during the 120-days before planting.     

Effects of nitrogen dioxide on selected soil processes

Nitrogen dioxide gas was rapidly absorbed by soil. After a 15 min incubation at 25°C, soil at a moisture content of 16% absorbed 99% of the NO₂ introduced into the gas-phase volume of a closed system. The presence of microorganisms hatl no influence on the rate of absorption of the gas by soil. The absorption of NO₂ by sandy clay loam soil was not an oxygen- or temperature-dependent process nor did it depend upon the moisture content of the soil. These physical factors acquired significance only in determining the initial rate of absorption of the gas and the rate at which NO₂ diffused through the soil. Exposure of soil to NO₂ resulted in substantial increases in the levels of NO _inf2 ^sup? N in the soil. Chemical oxidation of the NO _inf2 ^sup? N resulted in an increase in NO _inf3 ^sup? N levels. During a 14-day incubation, NO _inf2 ^sup? N concentrations in sterile soil exposed to an atmosphere containing 100 μg ml^?1 of NO₂ decreased from 190 μg g^?1 of soil to 105 μg g^?1 with an accompanying increase in NO _inf3 ^sup? N from 2 μg g^? ₁ to 63 μg g^?1 of soil. Nitrogen dioxide severely inhibited the growth of both aerobic and anaerobic asymbiotic N₂-fixing bacteria in soil. After a 48 h incubation at 25°C, soil aggregates exposed to an atmosphere containing 100 μg ml^?1 of NO₂ contained 88% and 98% fewer aerobic and anaerobic N₂-fixing bacteria, respectively. C₂H₂-reduction measurements showed that nitrogenase synthesis and activity in artificial soil aggregates amended with 2% glucose were inhibited by 20% and 48%, respectively, when exposed to atmospheric concentrations of 35 and 3.5 μg ml^?1 of NO₂, respectively.     

Effects of dredged spoils on selected microbial activities in lake column simulators

Four Lake Column Simulators were filled with hypolimnetic water from Lake Ontario and inoculated with algae and fish. Various microbial processes were monitored before and after the addition of dredged spoils. Loading rates of dredged spoils of 0.33, 3.3 or 33 g day^?1 were used for each of three columns and one column with no addition of dredged spoils was maintained as a control. The effects of dredged spoils on microbial activities were most pronounced at the highest loading rate. ³²P-PO₄ was rapidly removed from the control water but the presence of suspended particles from dredged material increased both biological assimilation and physical adsorption processes. The influence of dredged spoils on ³²P-PO₄ uptake was most pronounced immediately following loading. Dredged spoils also increased ¹⁴C-acetate mineralization in water and the effects were again more pronounced immediately following loading. A 3 fold increase in heterotrophic activity during a 1 h incubation at 20°C was observed following addition of a 33 g day^?1 loading. Dredged spoils increased the levels of N₂-fixation in the columns as indicated by the C₂H₂-reduction assay procedure. Suspended dredged material reduced light penetration and decreased primary production in the columns. The increased availability of dissolved and particulate organic and inorganic nutrients from continuous addition of dredged spoils was responsible for a general stimulation of microbial activities in the epilimnetic and hypolimnetic regions of the simulated lake columns.     

Conversion factor between acetylene reduction and nitrogen fixation in soil: Effect of water content and nitrogenase activity

The conversion factor between C₂H₂ reduction and N₂ fixation was studied in two soils. In one study small cores from (he two soils were drained to three water tensions: 0.20, 1.17 and 4.89 kPa. At each tension the N₂ase activity was measured with both O.1 aim ¹⁵N₂ and 0.1 aim C₂H₂. The conversion factor was different for the two soils. 1.0 and 3.1. respectively. The water content had no influence on the value of the conversion factor in this first study, in which the fixation corresponded to about 1mg N m^?2 day ^?1 at the depth 0–3 cm.In another study glucose was added to one of the soils to enhance the N₂ase activity. The activity was measured at 75 and 100% water saturation with both 0.9 atm ¹⁵N₂ and 0.1 atm C₂ H₂. At the lower water content the conversion factor was 2.6 and at water saturation the factor was 15.7. The fixation rates were high in this study. 98 mg N m^?2 day^?1 at the lower water content and 42 mg at water saturation.By theoretical calculations it was shown that the concentration of dissolved N₂ restricted the rate of fixation in the water-saturated samples of the second study, thus giving the high conversion factor. The critical level of N₂asc activity in water-saturated soil, above which the actual C₂H₂ to N₂-ratio will be higher than usual, was estimated to about 10mg Nm ²day_?1, under the experimental conditions used in these studies.     

Toxic potential of different types of sewage sludge as fertiliser in agriculture: ecotoxicological effects on aquatic,sediment and soil indicator species

Purpose

Treated and processed sewage sludges (biosolids) generated during the treatment of wastewater usually contain substantial concentrations of nutrients, especially phosphorus, which is essential for plant growth. Sewage sludge therefore can be used as an alternative fertiliser in agriculture. But since sewage sludge could also contain pollutants, analysis and ecotoxicological tests on affected soil and stream water organisms are necessary in order to guarantee its harmless use.

Materials and methods

Three test species were chosen to cover the environmental compartments, water, sediment and soil. The following test species and parameters were applied to evaluate the acute effects of three sewage sludge samples: Lemna minor (growth inhibition, discolouration and colony breakup), Gammarus fossarum (mortality, behaviour) and Eisenia fetida (avoidance behaviour). Chemical assessment included nutrients, organic pollutants and heavy metals.

Results and discussion

The assessment of a non-dewatered sludge (S1) sample resulted in an inhibition of growth of L. minor starting from 0.6 g total solid (TS)?l^?1 after 7 days (EC₅₀ 1.2 g TS l^?1). G. fossarum displayed significantly decreased movement activity at 0.5 and 1.2 g TS l^?1 sludge concentration during an exposure time of 2 days, leading to decreased survival after 4 days of exposure in 0.5 g TS l^?1 (LC₅₀ 0.5 g TS l^?1). After 2 days, E. fetida exhibited an increased avoidance behaviour of contaminated soil from 0.2 g TS kg^?1 sewage sludge (EC₅₀ 0.4 g TS kg^?1). The dewatered sludge samples (S2 and S3) had a lower toxic effect on the test organisms. G. fossarum was the most sensitive test species in the applied test setups. The realistic application amounts of the tested sewage sludge samples of approximately 6.0 g TS kg^?1 (maximum allowed application amount of sewage sludge) and approximately 3 g TS kg^?1 (maximum agronomical relevant application amount) in worst case studies are higher than the analysed EC₅₀/LC₅₀ values of S1 and of the LC₅₀ (G. fossarum) of S2 and S3.

Conclusions

All three tested sewage sludge samples have to be classified as toxic at high concentration levels under laboratory conditions. Realistic output quantities of S1 will negatively influence soil invertebrates and freshwater organisms (plants and crustacean), whereas the dewatered sludge samples will most likely not have any acute toxic effect on the test organisms in the field. Test with environmental samples should be conducted in order to support this hypothesis.

     

Acetylene inhibition of nitrous oxide reduction and measurement of denitrification and nitrogen fixation in soil

Reduction of N₂O in moist soil was inhibited completely by 10^?2 atm C₂H₂ and partially by 10^?5 atm C₂H₂. The effect of C₂H₄ was 10⁴ times less than that of C₂H₂. Denitrification of NO^?₃ occurred in anaerobically or aerobically incubated waterlogged soil and in anaerobic but not in aerobic moist soil. In the absence of C₂H₂ there was transient accumulation of N₂O. In the presence of C₂H₂ there was stoichiometric conversion of NO^?₃ to N₂O. Some kinetics of the reduction of N₂O and of NO^?₃ to N₂O are presented. Denitrification of 1 μg added NO^?₃-N.g^? could be measured within 1 h. Stoichiometries of production of N₂O from NO^?₂ and NO^?₃, respectively, and production of CO₂ attributable to denitrification were consistent with reported energy yields. Reduction of C₂H₂ to C₂H₄ occurred immediately following complete denitrification of added NO^?₃. The incubation of soil in the presence and in the absence of C₂H₂ thus permits assay of both denitrification and N₂ fixation and provides information on the mole fraction of N₂O in the products of denitrification.     

Combined biochar and nitrogen fertilizer reduces soil acidity and promotes nutrient use efficiency by soybean crop

Purpose

Soil acidification is universal in soybean-growing fields. The aim of our research was to evaluate the effects of soil additives (N fertilizers and biochar) on crop performance and soil quality with specific emphasis on ameliorating soil acidity.

Materials and methods

Four nitrogen treatments were applied as follows: no nitrogen (N0), urea (N1), potassium nitrate (N2), and ammonium sulfate (N3), each providing 30 kg N ha^?1. Half plot area of the N1, N2, and N3 treatments was also treated with biochar (19.5 t ha^?1) to form N-biochar treatments (N1C, N2C, N3C). Both bulk and rhizosphere soils were sampled separately for the following analyses: pH, exchangeable base cations (EBC), exchangeable acidity (EA), total inorganic N (IN), total N (TN), and microbial phospholipid fatty acids (PLFAs). Soybean biomass and nutrient contents were also determined. Correlation analysis was applied to analyze the relationships between soil chemical properties and soybean plant parameters.

Results and discussion

With N-biochar additions (N1C, N2C, N3C), soil chemical properties changed as follows: pH increased by 0.6–1.2 units, EBC, IN, and TN increased by 175–419, 38.5–54.7, and 136–452 mg kg^?1, respectively, and PLFAs increased by 23.6–40.9 nmol g^?1 compared to the N0 in the rhizosphere. Microbial PLFAs had positive correlations with soil pH; EBC; exchangeable K, Ca, Na, and Mg; TN; IN; NH₄ ⁺; and NO₃ ^? (r?=?0.66–0.84, p?<?0.01). There were negative correlations between PLFAs and EA or exchangeable Al (r?=??0.64, ?0.66, p?<?0.01), which indicated that the additives increased microbial biomass by providing a suitable environment with less acid stress and more nutrients. The additives increased soil NH₄ ⁺ and NO₃ ^? by promoting soil organic N mineralization and reducing NH₄ ⁺ and NO₃ ^? leaching. Moreover, the soybean seed biomass and the nutrient contents in seeds increased with N-biochar additions, especially in the N3C treatment.

Conclusions

N-biochar additions were effective in ameliorating soil acidity, which improved the microenvironment for more microbial survival. N-biochars influenced N transformations at the plant–soil interface by increasing organic N mineralization, reducing N leaching, and promoting N uptake by soybeans. The soil additive ammonium and biochar (N3C) were best in promoting soybean growth.

     

Structure-function relationship of vermicompost humic fractions for use in agriculture

Purpose

The use of humic substances (HS) in agriculture is beneficial and has positive environmental impacts. However, to optimize the use of HS possible links between their structural characteristics and bioactivity must be shown. The goal of this study is to evaluate the bioactivity of different humic fractions extracted from vermicompost (VC) in rice plants and to shed light to possible structure-function relationships.

Materials and methods

Humic-like fractions were obtained from cattle manure vermicompost processed by African nightcrawlers (Eudrilus eugeniae spp.). Humic-like acid fraction using only water as extractor (HLAw), HLA fraction extracted following the International Humic Substances Society (IHSS) recommended method, and the solid residue (humified residual (HR)) after extraction of HLA were characterized using complementary chemical, physic, and spectroscopic technics (elemental composition, UV-Vis and Fourier transform infrared spectroscopy (FTIR) spectroscopies, ¹³C-CP MAS NMR, and MEV). Biological activity of the three HS was conducted in growth chambers and measured in roots using WinRhizo Arabidopsis software. Principal component analysis (PCA) was used to find a grouping pattern between the structural variables evaluated and the obtained root parameters.

Results and discussion

Differences were found in elemental composition among HS with larger C/N ratio in HR than in HLA and HLAw. HLA and HLAw FTIR spectra showed carboxyl band at 1714.66 cm^?1 better resolved than in HR. Bands at 1642 cm^?1 (amide I) and 1510 cm^?1 (lignin), were better resolved in HLA. ¹³C-NMR showed the following order of aromaticity: HLA > HLAw > HR. For HLAw bioactivity, the structures C_Alkyl-H,R, C_C=O, and C_COO-H,R correlated with the number and growth of smaller root. The aromatic C_Ar-H,R, C_Ar-O,N, and aliphatic C_Alkyl-O,N, C_Alkyl-O, and C_Alkyl-di-O structures in HLA, correlated with larger roots growth. HR also stimulated root growth and development in rice plants.

Conclusions

Aliphatic and oxygenated structures in HLAw showed a relation with induction of initial root emissions, whereas the presence of aromatic compounds in HLA was related with root growth stimulation activity. Higher concentration of HLAw was necessary to produce an equivalent stimulus compared with HLA; it could indicate that, although both fractions showed similar types of structures in their composition, differences in the predominant structures may be determining different effects on the root.

     

Production and consumption of N2O during denitrification in subtropical soils of China

Purpose

Nitrous oxide (N₂O) production and reduction rates are dependent on the interactions with each other and it is therefore important to evaluate them within the context of simultaneously operating N₂O emission and reduction. The objective of this study was to quantify the simultaneously occurring N₂O emission and reduction across a range of subtropical soils in China, to gain a mechanistic understanding of potential N₂O dynamics under the denitrification condition and their important drivers, and to evaluate the potential role of the subtropical soils as either sources or sinks of N₂O through denitrification.

Materials and methods

Soils (45, from a range of different land uses and soil parent materials) were collected from the subtropical region of Jiangxi Province, China, and tested for their potential capacity for N₂O emission and N₂O reduction to N₂ during denitrification. N₂O emission and reduction were determined in a closed system under N₂ headspace after the soils were treated with 200?mg?kg^?1 NO ₃ ^? -N and incubation at 30?°C for 28?days. The soil physical and chemical properties, the temporal variations in headspace N₂O concentration, and NO ₃ ^? -N and NH ₄ ⁺ -N concentrations in the soil slurry were measured.

Results and discussion

Variations in N₂O concentration (N) over incubation time (t) were consistent with an equation in which average R ²?=?0.84?±?0.11 (p?<?0.05): $ N = A \times \left( {1 - \exp \left( { - {k_1} \times t} \right)} \right) - B \times \exp \left( {{k_2} \times t} \right) $ , where A is the total N₂O emission during the incubation, B is a constant, and k ₁ and k ₂ are the N₂O emission constant and reduction constants, respectively. The results of the simulation showed that k ₁ was greater than k ₂. The reduced amount of NO ₃ ^? -N in the first 7?days of incubation and the N₂O emission rate (the percentage of A value relative to the amount of NO ₃ ^? -N reduced during the 28-day incubation, R _n) were able to explain 82.9?% (p?<?0.01) of the variation in total N₂O emission (A) during the incubation for the soil samples studied, indicating that the total amount of N₂O emitted was determined predominately by denitrification capacity. Soil organic carbon content and soil nitrogen mineralization are the key factors that determine differences in the amounts of reduced NO ₃ ^? -N among the soil samples. The R _n value decreased with increasing k ₂ (p?<?0.01), indicating that soils with higher N₂O reduction capacity under these incubation conditions would emit less N₂O per unit of denitrified NO ₃ ^? -N than the other soils. Results are valuable in the evaluation of net N₂O emissions in the subtropical soils and the global N budget.

Conclusions

In a closed, anaerobic system, variations in N₂O concentration in the headspace over the incubation time were found to be compatible with a nonlinear equation. Soil organic carbon and the amount of NH ₄ ⁺ -N mineralized from the organic N during the first 7?days of incubation are the key factors that determine differences in the N₂O emission constant (k ₁), the N₂O reduction constant (k ₂), the total N₂O emission during the incubation (A) and the N₂O emission rate (R _n).     

Bacterivore nematodes stimulate soil gross N transformation rates depending on their species

We conducted a microcosm experiment with soil being sterilized, reinoculated with native microbial community and subsequently manipulated the bacterivorous nematodes, including three treatments: without (CK) or with introducing one species of the two bacterivores characterized with different body size but similar c-p (colonizer-persister) value (Rhabditis intermedia and Protorhabditis oxyuroides, accounted for 6 and 59% of bacterivores in initially undisturbed soil, respectively). We monitored the N₂O and CO₂ emissions, soil properties, and especially quantified gross N transformation rates using ¹⁵N tracing technique after the 50 days incubation. No significant differences were observed on soil NH₄ ⁺ and NO₃ ^? concentrations between the CK and two bacterivores, but this was not the case for gross N transformation rates. In comparison to CK, R. intermedia did not affect soil N transformation rates, while P. oxyuroides significantly increased the rates of mineralization of organic N to NH₄ ⁺, oxidation of NH₄ ⁺ to NO₃ ^?, immobilization of NO₃ ^? to organic N and dissimilatory NO₃ ^? reduction to NH₄ ⁺. Furthermore, the mean residence time of NH₄ ⁺ and NO₃ ^? pool was greatly lowered by P. oxyuroides, suggesting it stimulated soil N turnover. Such stimulatory effect was unrelated to the changes in abundance of bacteria and ammonia-oxidizing bacteria (AOB). In contrast to CK, only P. oxyuroides significantly promoted soil N₂O and CO₂ emissions. Noticeably, bacterivores increased the mineralization of recalcitrant organic N but decreased soil δ¹³C_-TOC and δ¹⁵N_-TN values, in particular for P. oxyuroides. Combining trait-based approach and isotope-based analysis showed high potential in moving forward to a mechanistic understanding of bacterivore-mediated N cycling.     

Extraction and speciation analysis of roxarsone and its metabolites from soils with different physicochemical properties

Purpose

The application of roxarsone (ROX), an arsenic-containing compound, as a feed additive in the animal production industry results in elevated soil levels of ROX and its metabolites, namely, monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), arsenate (As(V)), and arsenite (As(III)). This study was conducted to study the extraction and speciation analysis of ROX-related arsenicals in soils with different physicochemical properties and the possible effects of soil properties on the extraction of ROX and its metabolites.

Materials and methods

Analytical method based on high-performance liquid chromatography (HPLC)-inductively coupled plasma–mass spectrometry (ICP-MS) was employed to determine the concentrations of As(III), DMA, MMA, As(V), and ROX extracted by different extraction solvents from different soils spiked by arsenicals. Validity of the developed method was assessed by the recovery efficiencies of arsenic species in soil-dissolved matter solutions containing 20 μg As?·?L^?1 of each arsenic species. Effects of soil properties on the extraction of ROX and its metabolites were analyzed by Pearson’s correlation.

Results and discussion

Arsenic species were separated using gradient elution of water and 20 mmol?·?L^?1 (NH₄)₂HPO₄ + 20 mmol?·?L^?1 NH₄NO₃ + 5 % methanol (v/v) within 27 min. The linear ranges of all arsenicals were 0–200 μg As?·?L^?1 with R ²?>?0.9996. The developed method provided lower limits of detection for As(III), DMA, MMA, As(V), and ROX (0.80, 0.58, 0.35, 0.24, and 1.52 μg As?·?L^?1, respectively) and excellent recoveries (92.52–102.2 %) for all five species. Arsenic speciation was not altered by 0.1 mol?·?L^?1 NaH₂PO₄ + 0.1 mol?·?L^?1 H₃PO₄ (9:1, v/v), which offered better average extraction efficiencies for As(III), As(V), DMA, MMA, and ROX (32.49, 92.50, 78.24, 77.64, and 84.54 %, respectively). Extraction performance of arsenicals was influenced by soil properties, including pH, cation exchange capacity (CEC), total Fe, and amorphous Fe.

Conclusions

ROX and its metabolites from soils could be satisfactorily separated by the developed method for the studied arsenicals. To extract arsenic species from soils, 0.1 mol?·?L^?1 NaH₂PO₄ + 0.1 mol?·?L^?1 H₃PO₄ (9:1, v/v) was recommended. Extraction efficiencies of arsenicals were influenced more by solvent composition than soil physicochemical properties. The present study provides a valuable tool and useful information for determining the concentrations of ROX and its metabolites in contaminated soils.

     

The potential of using alternative pastures,forage crops and gibberellic acid to mitigate nitrous oxide emissions

Purpose

In grazed pastures, nitrous oxide (N₂O), a powerful greenhouse gas and an ozone depletion substance, is mostly emitted from animal excreta, particularly animal urine-N returned to the soil during grazing. We conducted a series of four field lysimeter and plot experiments to assess the potential of using gibberellic acid (GA) and/or alternative pastures or forage crops to mitigate N₂O emissions from outdoor dairy farming systems.

Materials and methods

Pasture and forage plants assessed in the experiments included Italian ryegrass (Lolium multiflorum L.), lucerne (Medicago sativa L.), diverse pastures (including plantain (Plantago lanceolata L.), chicory (Cichorium intybus L.), perennial ryegrass (Lolium perenne L.) and white clover (Trifolium repens L.)), fodder beet (Beta vulgaris L.), kale (Brassica oleracea L.), as well as the standard perennial ryegrass and white clover (RG/WC) pastures. N₂O was determined using a standard static chamber method in the field either on top of lysimeters or field plots.

Results and discussion

The results showed that the application of GA to urine-treated lysimeters with Italian ryegrass, lucerne or RG/WC pastures did not result in lower N₂O emissions. However, the use of diverse pastures which included plantain with a lower urine-N loading rate at about 500 kg N ha^?1 significantly decreased N₂O emissions by 46 % compared with standard RG/WC with a urine-N loading rate at 700 kg N ha^?1. However, when urine-N was applied at the same rates (at 500 or 700 kg N ha^?1), the N₂O emissions were similar between the diverse and the standard RG/WC pastures. This would indicate that it is the N-loading rate in the urine from the different pastures that determines the N₂O emissions from different pastures or forages, rather than the plants per se. The N₂O emissions from cow urine from fodder beet were 39 % lower than from kale with the same urine-N application rate (300 kg N ha^?1).

Conclusions

These results suggest that N₂O emissions can potentially be reduced by incorporating diverse pastures and fodder beet into the grazed pasture farm system. Further studies on possible mechanisms for the lower N₂O emissions from the different pastures or forages would be useful.

     

N<Subscript>2</Subscript>O production pathways relate to land use type in acidic soils in subtropical China

Purpose

Agricultural practises impact soil properties and N transformation rate, and have a greater effect on N₂O production pathways in agricultural soils compared with natural woodland soils. However, whether agricultural land use affects N₂O production pathways in acidic soils in subtropical regions remains unknown.

Materials and methods

In this study, we collected natural woodland soil (WD) and three types of agricultural soils, namely upland agricultural (UA), tea plantation (TP) and bamboo plantation (BP) soils. We performed paired ¹⁵N-tracing experiment to investigate the effects of land use types on N₂O production pathways in acidic soils in subtropical regions in China.

Results and discussion

The results revealed that heterotrophic nitrification is the dominant pathway of N₂O production in WD, accounting for 44.6 % of N₂O emissions, whereas heterotrophic nitrification contributed less than 2.7 % in all three agricultural soils, due to a lower organic C content and soil C/N ratio. In contrast, denitrification dominated N₂O production in agricultural soils, accounting for 54.5, 72.8 and 77.1 % in UA, TP and BP, respectively. Nitrate (NO₃ ^?) predominantly affected the contribution from denitrification in soils under different land use types. Autotrophic nitrification increased after the conversion of woodland to agricultural lands, peaking at 42.8 % in UA compared with only 21.5 % in WD, and was positively correlated with soil pH. Our data suggest that pH plays a great role in controlling N₂O emissions through autotrophic nitrification following conversion of woodland to agricultural lands.

Conclusions

Our results demonstrate the variability in N₂O production pathways in soils of different land use types. Soil pH, the quantity and quality of organic C and NO₃ ^? content primarily determined N₂O emissions. These results will likely assist modelling and mitigation of N₂O emissions from different land use types in subtropical acidic soils in China and elsewhere.

     

Nitrogen fixation by intact grass-soil cores using 15N2 and acetylene reduction

To determine N₂ fixation by intact grass-soil cores, samples were collected from 25 sites in central Texas during the summer. Three cores (32 cm² each) were extracted immediately adjacent to one another from single grass clumps or sods. Two of these cores were incubated under 10% C₂H₂ in air and the third core was incubated for 12 h in an atmosphere with 10% ¹⁵N₂ enrichment. Following incubation with ¹⁵N₂ the same core was assayed for rate of C₂H₂ reduction (AR). Rates of AR were generally low and quite variable (0–7.6 μmol C₂H₄ core^?1 day^?1). ¹⁵N₂ was incorporated into root and shoot tissues within 12–24 h. Extrapolated values of N₂ fixation based on ¹⁵N₂ incorporation ranged from 0 to 20 kg N ha^?1100 day^?1. The ratio of C₂H₂ reduced (μ mol C₂H₄ core^?1 day^?1) to N₂ fixed (μ mol N₂ fixed core^?1 day^?1) was highly variable ranging from 0 to 12. This study confirmed that N₂ is fixed in the rhizosphere of grasses grown in Texas through the use of ¹⁵N₂ and demonstrated that incorporation of fixed N into shoots was relatively rapid.