Can differences in 15N natural abundance be used to quantify the transfer of nitrogen from legumes to neighbouring non-legume plant species?

Natural variations in the stable isotope ¹⁵N are often exploited in studies of N cycling in ecosystems. Lower ¹⁵N natural abundance in non-legume plants growing in association with legumes, compared with the non-legume grown alone in pure stands have been observed in cropping, forage, and agroforestry systems. Such observations have frequently been attributed to the transfer of biologically-fixed nitrogen (N) from the legume to the companion non-legume, and various methodologies have been employed to calculate the extent of the N transfer. While some of these ¹⁵N natural abundance-based estimates of N transfer were within the range previously reported using equivalent ¹⁵N-enriched techniques (<20% of non-legume plant N and <10 kg N ha⁻¹ derived from fixed N contributed by neighbouring legumes), many of the values obtained using natural abundance were much higher (30%–83% of the non-legume N derived from fixed N representing up to 30–40 kg N ha⁻¹) than generally measured by ¹⁵N-enriched methods; with even greater estimates being determined where data were available to allow N transfer to be re-calculated on the basis of total legume N rather than fixed N (42% to >100%, and up to 110 kg N ha⁻¹ per year). This review raises concerns about the assumptions behind the natural abundance approach, and provides some alternative interpretations for the observed differences in natural ¹⁵N abundance between plants grown in the presence and absence of legumes. It was concluded that simple comparative measures of non-legume δ¹⁵N alone cannot provide a quantitative estimate of N transfer between plant species if the dominant source and the isotopic identity of the transferred N cannot be validated, and if the extent of any isotopic fractionation associated with relevant N transformations occurring during transfer cannot be defined. To date this information is not forthcoming. There is a need to greatly improve our understanding of the transfer processes before the real value of the δ¹⁵N technology can be realized. In the first instance this will primarily be achieved by carefully executed experiments under controlled conditions, and in the field, employing both ¹⁵N natural abundance and enrichment approaches so estimates of transfer can be compared, and the data interrogated using modelling approaches to explore isotopic fractionation.     

陕北不同土地利用类型下土壤无机磷形态分布及有效性研究

采用蒋柏藩-顾益初无机磷分级方法,对黄土高原两种农田土壤剖面(0～100 cm)中无机磷形态分布特征进行了研究.结果表明:两种农田土壤剖面(0～100 cm)中无机磷形态占全磷的90.95%;无机磷又以Ca-P为主,所占比例在87.86%～92.55%之间;糜子地土壤剖面中无机磷总量、Ca2-P、Ca8-P、Al-P和...     

Changes in Cd,Cu, Ni,Pb and Zn Fractionation and Liberation Due to Mussel Shell Amendment on a Mine Soil

Mining activities are related to relevant environmental pollution issues that should be controlled. We used sequential extractions to fractionate Cd, Cu, Ni, Pb and Zn retained on unamended or mussel shell‐amended mine soil samples, all of them treated with a mixture of the five heavy metals (total metal concentration of 1·57 mmol L⁻¹), after 1, 7 and 30 days of incubation. In addition, we used the stirred flow chamber technique to study the release of each of the five heavy metals from these different unamended and shell‐amended soil samples. The results indicate that the shell amendment caused a decrease in the most soluble fraction, while increasing the most recalcitrant (least mobile) fraction. With equivalent implications, the stirred flow chamber experiments showed that mussel shell amendment was associated to a decrease in heavy metal release and increased retention. The highest mussel shell dose and incubation time caused the most relevant changes in pH values and thus in metal retention, also indicating the importance of pH modifications in the mechanism of retention acting in the amended samples. In view of these results, the use of mussel shell amendment can be encouraged to increase heavy metal retention in acid mine soils, in order to minimise risks of environmental pollution. Copyright © 2016 John Wiley & Sons, Ltd.     

Phosphorus content in five representative landscape units of the Lomas de Arequipa (Atacama Desert-Peru)

Phosphorus forms and content were studied in soils of the Lomas de Arequipa (Atacama desert, Peru) using a fractionation method. These Lomas are small hills periodically submitted to the El Niño-Southern Oscillation (ENSO) which causes heavy rainfall. Sample soils were randomly selected in five landscape types characterized by vegetation: cactaceae (Cac), cactaceae and herbaceous (CacHerb), shrubs (Shr), trees with cover < 60% (Tree) and shrubs or trees with cover > 60%) (ShrTree). All the soils were strongly acidic and classified as loamy sand, sandy loam or silt loam. Organic carbon content was under 1% in Cac or CacHerb, then increased strongly in ShrTree (6.50%). Considering phosphorus, all the forms (labile as well resistant forms) increased markedly from Cac soils to ShrTree soils. In all the soils, the labile forms (Resin-P: range 45–105 μg g^− 1; NaHCO3-Pi: 23–123 μg g^− 1; or NaHCO3-Po: 10–122 μg g^− 1) were very high. These high phosphorus contents were attributed to the specific climatic conditions of the Lomas that feature a long period of vegetation dormancy (very dry period) and a short period of growth, following ENSO-associated precipitation. We suggested that during the dry period, plant decay and microbial cells death lead to release and accumulation of labile P in the soil, the rainfall wetting the soil, permitting vegetation growth. In this respect, the Lomas climatic conditions contribute to soil fertility, especially as labile forms of phosphorus are chiefly concerned.     

Harmonisation, standardisation for soil and sediment fractionation studies, and usefulness in assessment and risk management

Single and sequential extraction procedures are often used to study the environmental fate of trace elements from soils and sediments. Although these schemes are quite popular, they are prone to various sources of errors, which require them to be thoroughly controlled. Owing to their operationally-defined character, the only way to compare analytical data is to stick closely to procedures that have been collaboratively studied and written in the form of an operating protocol. Groups of scientists have hence developed ‘harmonised’ procedures that are used as a reference for fractionation studies. A higher level is the formal standardisation of the procedures by official standardisation bodies, which also fulfils the need for achieving the comparability of data obtained by different laboratories from different countries. This paper discusses aspects of harmonisation (consensus obtained by groups of scientists) and standardisation (adoption of standards as mandated by an international organisation) as applied to single and extraction schemes for trace element fractionation studies, including quality control aspects.     

Natural N abundances of inorganic nitrogen in soil treated with fertilizer and compost under changing soil moisture regimes

This study was conducted to examine whether the applications of N-inputs (compost and fertilizer) having different N isotopic compositions (δ¹⁵N) produce isotopically different inorganic-N and to investigate the effect of soil moisture regimes on the temporal variations in the δ¹⁵N of inorganic-N in soils. To do so, the temporal variations in the concentrations and the δ¹⁵N of NH₄⁺ and NO₃⁻ in soils treated with two levels (0 and 150 mg N kg⁻¹) of ammonium sulfate (δ¹⁵N=−2.3‰) and compost (+13.9‰) during a 10-week incubation were compared by changing soil moisture regime after 6 weeks either from saturated to unsaturated conditions or vice versa. Another incubation study using ¹⁵N-labeled ammonium sulfate (3.05 ¹⁵N atom%) was conducted to estimate the rates of nitrification and denitrification with a numerical model FLUAZ. The δ¹⁵N values of NH₄⁺ and NO₃⁻ were greatly affected by the availability of substrate for each of the nitrification and denitrification processes and the soil moisture status that affects the relative predominance between the two processes. Under saturated conditions for 6 weeks, the δ¹⁵N of NH₄⁺ in soils treated with fertilizer progressively increased from +2.9‰ at 0.5 week to +18.9‰ at 6 weeks due to nitrification. During the same period, NO₃⁻ concentrations were consistently low and the corresponding δ¹⁵N increased from +16.3 to +39.2‰ through denitrification. Under subsequent water-unsaturated conditions, the NO₃⁻ concentrations increased through nitrification, which resulted in the decrease in the δ¹⁵N of NO₃⁻. In soils, which were unsaturated for the first 6-weeks incubation, the δ¹⁵N of NH₄⁺ increased sharply at 0.5 week due to fast nitrification. On the other hand, the δ¹⁵N of NO₃⁻ showed the lowest value at 0.5 week due to incomplete nitrification, but after a subsequence increase, they remained stable while nitrification and denitrification were negligible between 1 and 6 weeks. Changing to saturated conditions after the initial 6-weeks incubation, however, increased the δ¹⁵N of NO₃⁻ progressively with a concurrent decrease in NO₃⁻ concentration through denitrification. The differences in δ¹⁵N of NO⁻₃ between compost and fertilizer treatments were consistent throughout the incubation period. The δ¹⁵N of NO₃⁻ increased with the addition of compost (range: +13.0 to +35.4‰), but decreased with the addition of fertilizer (−10.8 to +11.4‰), thus resulting in intermediate values in soils receiving both fertilizer and compost (−3.5 to +20.3‰). Therefore, such differences in δ¹⁵N of NO₃⁻ observed in this study suggest a possibility that the δ¹⁵N of upland-grown plants receiving compost would be higher than those treated with fertilizer because NO₃⁻ is the most abundant N for plant uptake in upland soils.     

Recovering decomposing plant residues from the particulate soil organic matter fraction: size versus density separation

 A detailed size separation of particulate organic matter (POM) from soils amended with straw from Hordeum vulgare or Vicia sativa revealed that the loss of C during the first 56 days of incubation mainly occurred from particles >2,000 μm, without a concomitant reduction in the size of these large particles. Preliminary studies of POM from non-amended soil had shown that the stable heavy (>1.4 g cm^–3) POM fraction was mainly (>80%) composed of particles <400 μm, whereas the light fraction was dominated by larger particles (>80%). Therefore we decided to compare the POM <1.4 g cm³ with POM >400 μm. There was a very close relationship between POM>400 μm and POM <1.4 g cm^–3 with regard to amounts of C and N, as well as the appearance of these fractions under the microscope. Similarly there was a close relationship between changes in the C content of the POM fractions and the CO₂ respired, and this was also the case when comparing changes in POM-N with net N mineralization. This indicated that the biological activity during decomposition was actually localized in the POM. Due to the lighter workload and lower expenditure for reagents in connection with size separation of POM, we recommend the size separation procedure in connection with studies of residue decomposition in arable systems. Received: 23 May 2000     

Bioassay-directed Fractionation of Organic Extracts of Marine Surface Sediments from the North and Baltic Sea - Part II: Results of the biotest battery and development of a biotest index (8 pp)

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Part I: Determination and identification of organic pollutants Part II: Results of the biotest battery and development of a biotest index

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Preamble. This series of two papers presents the results of an interdisciplinary research project (ISIS) dealing with bioassay-directed fractionation of marine sediment extracts. Part I presents the extraction and fractionation procedure as well as the results of chemical analysis, including non-target analysis of sediments. Part II describes the results of the biotest battery in relation to chemicals possibly causing parts of the observed effects. A biotest index is used to compare the toxicities of the samples.

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AUTHORS / AFFILIATIONS Ninja Reineke (3), Werner Wosniok (4), Dirk Danischewski (1), Heinrich Hühnerfuss (3), Angelika Kinder (5), Arne Sierts-Herrmann (5), Norbert Theobald (2), Hans-Heinrich Vahl (6), Michael Vobach (1), Johannes Westendorf (6) and Hans Steinhart (5).

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(1) Federal Research Centre for Fisheries, Institute for Fishery Ecology, Palmaille 9, 22767 Hamburg, Germany (2) Federal Maritime and Hydrographic Agency, Bernhard-Nochtstr. 78, 20359 Hamburg, Germany (3) University of Hamburg, Institute for Organic Chemistry, Martin-Luther-King-Platz 6, 20146 Hamburg, Germany (4) University of Bremen, Institute of Statistics, Bibliothekstr. 1, 28334 Bremen, Germany (5) University of Hamburg, Institute for Food Chemistry, Grindelallee 117, 20146 Hamburg, Germany (6) University of Hamburg, University Hospital Hamburg-Eppendorf, Department for Toxicology, Vogt-Kölln-Str. 30, 22527 Hamburg, Germany (7) Eurofins Wiertz-Eggert-Jörissen, Stenzelring 14b, 21107 Hamburg, Germany

Goal, Scope and Background

The ecological relevance of contaminants in mixtures is difficult to assess, because of possible interactions and due to lacking toxicity data for many substances present in environmental samples. Marine sediment extracts, which contain a mixture of environmental contaminants in low concentrations, were the object of this study. The extracts were investigated with a set of different biotests in order to identify the compound or the substance class responsible for the toxicity. For this goal, a combination of biotests, biotest-directed fractionation and chemical analysis has been applied. Further on, a strategy for the development of a biotest index to describe the toxicity of the fractions without a prior ranking of the test results is proposed. This article (Part II) focuses on the biological results of the approach.

Methods

The toxicological potential of organic extracts of sediments from the North Sea and the Baltic Sea was analyzed in a bioassay-directed fractionation procedure with a set of biotests: luciferase reporter gene assays on hormone receptor and Ah receptor, arabinose resistance test, fish embryo test (Danio rerio), comet assay, acetylcholinesterase inhibition test, heat-shock protein 70 induction, oxidative stress and luminescence inhibition test (Vibrio fischeri). The test results provided the basis for the calculation of a biotest index by factor analysis to compare the toxicity of the samples and fractions.

Results and Discussion

Results of 11 biotests on different fractionation levels of the samples were described and discussed with regard to the occurrence of contaminants and their toxic potentials. Polychlorinated biphenyls, polycyclic aromatic hydrocarbons, quinones, brominated indoles and brominated phenols were in the focus of interest. A biotest index was constructed to compare the toxic responses in the samples and to group the biotest results.

Conclusion

The procedure presented in this study is well suited for bioassay-directed fractionation of marine sediment extracts. However, in relatively low contaminated samples, high enrichment factors and sufficient fractionation is necessary to allow identification of low concentrations of contaminants which is required to link effects and possible causes. In the present case, the relation between substances and effects was difficult to uncover due to relatively low concentrations of pollutants compared to the biogenic matrix and to the remaining complexity of the fractions. The results, with respect to the brominated phenols and indoles in the samples, highlight the successful use of bioassay directed fractionation in the case of high concentrations and high toxicity.

Recommendation and Outlook

In general, it has been shown that a marine risk assessment requires focusing on the input of diffuse sources and taking into account the fact of mixture toxicity. Effects resulting from biogenic substances will make the assessment of the influence of anthropogenic substances even more difficult.     

Soil organic matter fractionation as a tool for predicting nitrogen mineralization in silty arable soils

After decades of searching for a practical method to estimate the N mineralization capacity of soil, there is still no consistent methodology. Indeed it is important to have practical methods to estimate soil nitrogen release for plant uptake and that should be appropriate, less time consuming, and cost effective for farmers. We fractionated soil organic matter (SOM) to assess different fractions of SOM as predictors for net N mineralization measured from repacked (disturbed) and intact (undisturbed) soil cores in 14 weeks of laboratory incubations. A soil set consisting of surface soil from 18 cereal and root‐cropped arable fields was physically fractionated into coarse and fine free particulate OM (coarse fPOM and fine fPOM), intra‐microaggregate particulate OM (iPOM) and silt and clay sized OM. The silt and clay sized OM was further chemically fractionated by oxidation with 6% NaOCl to isolate an oxidation‐resistant OM fraction, followed by extraction of mineral bound OM with 10% HF (HF‐res OM). Stepwise multiple linear regression yielded a significant relationship between the annual N mineralization (kg N/ha) from undisturbed soil and coarse fPOM N (kg N/ha), silt and clay N (kg N/ha) and its C:N ratio (R² = 0.80; P < 0.01). The relative annual N mineralization (% of soil N) from disturbed soils was related to coarse fPOM N, HF‐res OC (% of soil organic carbon) and its C:N ratio (R² = 0.83; P < 0.01). Physical fractions of SOM were thus found to be the most useful predictors for estimating the annual N mineralization rate of undisturbed soils. However, the bioavailability of physical fractions was changed due to the disturbance of soil. For disturbed soils, a presumed stable chemical SOM fraction was found to be a relevant predictor indicating that this fraction still contains bio‐available N. The latter prompted a revision in our reasoning behind selective oxidation and extraction as tools for characterizing soil organic N quality with respect to N availability. Nonetheless, the present study also underscores the potential of a combined physical and chemical fractionation procedure for isolating and quantifying N fractions which preferentially contribute to bulk soil N mineralization. The N content or C:N ratio of such fractions may be used to predict N mineralization in arable soils.     

Does returning sites of historic peri‐urban waste disposal to vegetable production pose a risk to human health? – A case study near Manchester,UK

Urban waste disposal occurred on fenland to the west of Manchester, England, between 1900 and 1964. The reclaimed fenland, Chat Moss, is now used for mixed arable farming. A total of 1.92 Mt of waste including privy midden, street sweepings, clinkers and slaughterhouse refuse was incorporated into the moss resulting in a modified topsoil with raised pH and reduced organic matter content compared with the subsoil. Elevated levels of potentially toxic elements (PTEs) are observed in the topsoil beyond the typical depth of atmospheric contamination; Cd and As concentrations exceed soil guideline values (SGVs) at 1.8 and 43 mg/kg, respectively. Sequential extraction indicates that waste‐derived Pb, Zn and Ni remain predominantly in the residual fraction, whereas Cu was mainly organically bound. Arsenic was predominately found in oxide and organic matter fractions with Cd in carbonate, oxide, organic matter and residual fractions. Pot trials indicated limited uptake of PTEs by vegetables grown on the waste‐amended soil, with the exception of Cd uptake by lettuce (0.22 mg/kg FW) and Pb uptake by radish (0.16 mg/kg FW), which exceeded current EU limits of 0.2 and 0.1 mg/kg FW, respectively. Hazard quotients (HQs) identified no risks to adults from consumption of vegetables grown in these soils with the exception of lettuce consumption with a HQ of 1.4. Risks to children were slightly greater with HQs >1 for Cd in lettuce, spinach, carrots and onion, As in lettuce, parsley and onion and for Zn in spinach.