Environmental fate and effects of Bacillus thuringiensis (Bt) proteins from transgenic crops: a review

This paper reviews the scientific literature addressing the environmental fate and nontarget effects of the Cry protein toxins from Bacillus thuringiensis (Bt), specifically resulting from their expression in transgenic crops. Published literature on analytical methodologies for the detection and quantification of the Cry proteins in environmental matrices is also reviewed, with discussion of the adequacy of the techniques for determining the persistence and mobility of the Bt proteins. In general, assessment of the nontarget effects of Bt protein toxins indicates that there is a low level of hazard to most groups of nontarget organisms, although some investigations are of limited ecological relevance. Some published reports on the persistence of the proteins in soil show short half-lives, whereas others show low-level residues lasting for many months. Improvements in analytical methods will allow a more complete understanding of the fate and significance of Bt proteins in the environment.     

A Chronic Plant Test for the Assessment of Contaminated Soils. Part 2: Testing of contaminated soils (10 pp)

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PREAMBLE According to the German Federal Soil Protection Act (BBodSchG 1998), the habitat function of soils must be protected. Despite the fact that in the Federal Soil Protection Ordinance (BBodSchV 1999) it has not been established how this goal can be reached reliably, it is clear that such a biological function can only adequately be assessed using biological test methods. This is especially true when a soil is contaminated by a mixture of often unknown chemicals. In such a case the use of chemical analysis aiming at a small range of known substances is not sufficient and must therefore be supplemented by biological methods. For this reason, several standardised test methods are available (e.g. using earthworms, collembolans or plants; Römbke and Knacker 2003; ISO 2003). Since acute tests are usually not sensitive enough for the assessment of potentially contaminated soils (e.g. Hund-Rinke et al. 2002), chronic tests like the earthworm reproduction tests (ISO 1998) are recommended for this purpose.

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A chronic plant test for the determination of phytotoxicity was missing until quite recently. The term phytotoxicity is understood here as the capacity of a compound or a contaminated soil to cause temporary or long-lasting damage to plants (EPPO 1997). Therefore, the German Ministry for Education and Research sponsored a project (1997 – 1999) in which – based on existing standardised methods – such a chronic plant laboratory test was developed and partly validated (Kalsch and Römbke 2000). The new test can be used for the evaluation of single chemicals (see Part 1 of this mini-series) as well as for the assessment of contaminated or remediated soils (see Part 2 of this mini-series).

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ABSTRACT Background and Scope. A new chronic plant test system which is based on experiences with various acute plant tests (e.g. published by OECD or ISO) and existing North American Plant-Life-Cycle Bioassays was standardised in a project sponsored by the German government. Characteristic properties of the test system, which can be performed either with Brassica rapa (turnip rape) or Avena sativa (oat), are described in Part I of this mini-series.

Methods

This new test was used to assess the effects of natural soil samples contaminated with TNT (2,4,6-Trinitrotoluene) or PAHs (poly-aromatic hydrocarbons). The soils were tested after taken from the field as well as after being remediated. Different control and reference soils were used to evaluate the test results. In addition, they were compared with the results of tests in which either TNT or Pyrene was spiked to field and standard soils (see Part I of this mini-series).

Results

All contaminated soils showed clear effects in the Chronic Plant Test (usually B. rapa was more sensitive than A. sativa). LUFA 2.2 standard soil and OECD artificial soil are well-suited as control and mixture substrates, while reference soils collected at uncontaminated sites were several times phytotoxic. In most of the latter cases, soil properties could be identified as the main cause of these effects (e.g. the pH value). While in general the sensitivity of the reproduction and biomass endpoints did not differ much, it is recommended to measure different endpoints (i.e. biomass and reproduction) due to the different mode-of-action of contaminants. In the case of TNT a good agreement between the results of single chemical tests and tests with TNT-contaminated soils was found (due to the minimal effects of Pyrene the same statement is not possible for PAHs).

Outlook

The results of this work have been used to prepare a draft test guideline, which has recently been standardised by the International Organisation for Standardisation (ISO). Based on these experiences, recommendations are given how this chronic test system could be used as part of a test battery for the assessment of contaminated soils.     

Collembola in ecotoxicology—Any news or just boring routine?

Despite the uncontested significance of soils for human nutrition and drinking water quality, the majority of ecotoxicological testing is confined to aquatic test systems. Among the standardised tests for soils, the reproduction test with the springtail Folsomia candida is among the most widely used ones. First steps towards its standardisation were undertaken in the late 1980s. Here we review major advances that have been made since then, with respect to mechanistic, pragmatic and ecological aspects. Specifically we address the ecological relevance of any modifications of the standardised tests. We introduce a miniaturised version of the reproduction test which allows reducing the amount of soil per test unit to one third and the number of synchronised individuals to 40% as compared to the standard test. In addition, we developed an assay using Collembola eggs instead of synchronised adults. First results of a three-species test indicate that the presence of other species may affect choice behaviour. We point out a potential biased view of existing ecotoxicological data with Collembola due to the fact that most results refer to metal contamination. Finally, recommendations for future research are given, with special reference to avoidance and microcosm tests involving Collembola.     

Technical experiences made with the litter bag test as required for the risk assessment of plant protection products in soil

Background, aim, and scope  The aim of this review was to assess the degree to which recent litter bag studies performed between 2002 and 2004 have followed the recommendations laid down in the ‘Effects of plant protection products on functional endpoints in soil (EPFES)’ guidance document and the Organisation for Economic Co-Operation and Development (OECD) Guidance Document on the Breakdown of Organic Matter in Litter bags (OECD, Guidance document on the breakdown of organic matter in litter bags, OECD series on testing and assessment, no 56, 36 pp, 2006), which can be required under European Union (EU) Directive 91/414/EEC (EU, Off J Eur Union, L230:1–32, 1991) for assessing the effect of plant protection products on non-target soil macro-organisms which contribute to the breakdown of organic matter. Materials and methods  The current review covers the analysis of key parameters of the litter bag method of a total of 29 litter bag studies conducted by six European Crop Protection Association companies. Results  The two validity criteria [i.e. 60% mass loss at the end of the study in the control and a maximum coefficient of variation of 40% for mass loss in the control plots (n = 6) during the first 6 months of the test] were met in all of the studies reviewed. In over half of the studies, more than 60% mass loss was reached in the control litter bags after about 6 months. Statistically significant effects on organic matter breakdown due to plant protection products were repeatedly observed during the first few months of the study conduct, including effects >10% up to >25% compared to the control. Discussion  The survey of 29 study reports—performed from May 2002 to May 2004—showed that litter bag studies are generally being performed following the ‘EPFES’ guidance document and the OECD guidance document no. 56. Transient, statistically significant effects >10% up to >25% compared to the control were determined repeatedly during the first few months of the study conduct, thus clearly indicating the sensitivity of the litter bag test system. Conclusions and perspectives  The litter bag study design is a valuable test system for the risk assessment of plant protection products on non-target soil macro-organisms which contribute to the breakdown of organic matter under EU Directive 91/414/EEC (EU, Off J Eur Union, L230:1–32, 1991). In general, it can be concluded that the litter bag test, as proposed by EPFES and the OECD guidance document, provides an integrative answer on the potential effects of plant protection products on organic matter breakdown in soil and can be continued to be used if the studies follow the recommendations as presented in the guidance documents closely.     

Tackling the heterogeneity of soils in ecotoxicological testing an euro-soil based approach

Goal, Scope and Background  In terrestrial ecotoxicology, standardised test methods using plants, earthworms and insects are available for the evaluation of effects induced by heavy metals, organic chemicals and, in particular, pesticides. Currently, these tests are performed either by using so-called Artificial Soils or (more or less) arbitrarily selected natural soils. Consequently, the test results depend not only on the intrinsic physico-chemical properties of the test chemical, but are also significantly influenced by the variable properties of the chosen soil In order to standardise the. test conditions and, at the same time, to relate the test results to representative soil types within the EU, it is proposed to modify the EURO-Soil concept for testing purposes. Main Features  The EURO-Soil concept, i.e. the selection of a limited number of soils that are representative for Europe, was originally developed for the performance of standardised environmental fate tests. Despite many problems in detail, soils from six sites all over Europe were identified that cover a wide range of soil properties (e.g. texture, pH, organic matter content) and, therefore, very different conditions concerning the bioavailability and, in turn, the effects of chemicals. Obviously, the routine use of EURO-Soils as a control or test substrate would require large amounts of soil. Therefore, it is proposed to modify this concept in such a way that all soils similar to one of the six EUROSoils can be used for ecotoxicological tests. Results and Discussion  It is assumed that the six EURO-Soils are representative for wide areas of the European Union, but at the same time it is neglected that some soils typical for, e.g. Northern Scandinavia, have to be identified in the future. All soils having similar properties (i.e. texture, pH, C/N ratio, and organic matter content) as one of the original EURO-Soils are called SIM-Soils. In this contribution, ‘ranges’ are proposed for four main properties and the six EUROSoils, thus allowing the identification of the SIM-Soils. However, since these properties cover a continuum, soils cannot be classified easily into a small number of classes; expert knowledge is required in order to decide whether a natural soil belongs to a certain SIM-Soil class or not. In the long run, this classification must take biological parameters like their suitability for standard test organisms into account as well. Recommendation and Outlook  The soils selected so far (at least one for each EURO-Soil) are actually tested using different biological test methods. Further tests are necessary in order to decide which tests can be done in which soil and whether new test systems, e.g. covering acid soils, have to be developed. However, it is already clear that the standard test species differ distinctly regarding their sensitivity towards soil properties. It is recommended to use the SIMSoils in order to provide the authorities with more field-relevant, data when assessing chemicals in the terrestrial environment.     

Stabilization of organic matter in temperate soils: mechanisms and their relevance under different soil conditions – a review

Mechanisms for C stabilization in soils have received much interest recently due to their relevance in the global C cycle. Here we review the mechanisms that are currently, but often contradictorily or inconsistently, considered to contribute to organic matter (OM) protection against decomposition in temperate soils: (i) selective preservation due to recalcitrance of OM, including plant litter, rhizodeposits, microbial products, humic polymers, and charred OM; (ii) spatial inaccessibility of OM against decomposer organisms due to occlusion, intercalation, hydrophobicity and encapsulation; and (iii) stabilization by interaction with mineral surfaces (Fe‐, Al‐, Mn‐oxides, phyllosilicates) and metal ions. Our goal is to assess the relevance of these mechanisms to the formation of soil OM during different stages of decomposition and under different soil conditions. The view that OM stabilization is dominated by the selective preservation of recalcitrant organic components that accumulate in proportion to their chemical properties can no longer be accepted. In contrast, our analysis of mechanisms shows that: (i) the soil biotic community is able to disintegrate any OM of natural origin; (ii) molecular recalcitrance of OM is relative, rather than absolute; (iii) recalcitrance is only important during early decomposition and in active surface soils; while (iv) during late decomposition and in the subsoil, the relevance of spatial inaccessibility and organo‐mineral interactions for SOM stabilization increases. We conclude that major difficulties in the understanding and prediction of SOM dynamics originate from the simultaneous operation of several mechanisms. We discuss knowledge gaps and promising directions of future research.     

Antioxidative stress activity of oligophosphopeptides derived from hen egg yolk phosvitin in Caco-2 cells

The protective effects of hen egg yolk phosvitin phosphopeptides (PPPs) against hydrogen peroxide (H2O2)-induced oxidative stress were evaluated in an in vitro assay using human intestinal epithelial cells. Caco-2 cells were stimulated with 1 mM H2O2 for 6 h, and the secretion of IL-8, a proinflammatory mediator, was determined by ELISA as a biomarker of oxidative stress. The inhibition of H2O2-induced IL-8 secretion from Caco-2 cells was observed by pretreatment for 2 h with PPPs, but not with phosvitin. PPPs also suppressed the formation of malondialdehyde in H2O2-treated Caco-2 cells. Furthermore, intracellular glutathione levels and glutathione reductase activity were elevated by the addition of PPPs. The protective effects of PPPs against H2O2-induced oxidative stress were almost the same as that of glutathione, and PPPs with a high content of phosphorus exhibited higher protective activity than PPPs without phosphorus; however, phosphoserine itself did not show any significant antioxidative stress activity. These findings suggest that oligophosphopeptides from hen egg yolk phosvitin possess novel antioxidative activity against oxidative stress in intestinal epithelial cells and that phosphorus and peptide structure seem to have a key role in the activity.     

Microbial respiration activities related to sequentially separated, particulate and water-soluble organic matter fractions from arable and forest topsoils

This study aimed to reveal differences in the relevance of particulate as well as water-soluble organic matter (OM) fractions from topsoils to the easily biodegradable soil organic matter (SOM). We selected eight paired sites with quite different soil types and soil properties. For each of these sites, we took samples from adjacent arable and forest topsoils. Physically uncomplexed, macro-, and micro-aggregate-occluded organic particle, as well as water-soluble OM fractions were sequentially separated by a combination of electrostatic attraction, ultrasonic treatment, density separation, sieving, and water extraction. The easily biodegradable SOM of the topsoil samples was determined by measuring microbial respiration during a short-term incubation experiment (OC_R). The organic carbon (OC) contents separated by i) the physically uncomplexed water-soluble OM, ii) the macro-, and iii) the micro-aggregate-occluded organic particle as well as water-soluble OM fractions were significantly correlated with OC_R. The correlation coefficients vary between 0.54 and 0.65 suggesting differences in the relevance of these OM fractions to the easily biodegradable SOM. The strongest correlation to OC_R was detected for the OC content separated by the physically uncomplexed water-soluble OM indicating the most distinct relation to the easily biodegradable SOM. This was found to be independent from land use or soil properties.     

Soil health – What should the doctor order?

The concept of soil health has been extensively reviewed in the scientific literature, but there is only patchy and inconsistent information available to farmers and growers who are concerned about the declining condition of their soils and are looking for appropriate test methods and management interventions to help reverse it. Although there are well‐established laboratory methods for soil chemical analysis, and a range of laboratory and field methods for measuring soil physical properties, only now are methods starting to emerge for soil biological analysis. This study provides an overview of the methods that are currently available commercially (or are close to commercialization) for farmers and growers in the UK. We examine the science underpinning the methods, the value of the information provided and how farmers and advisors can use results from such assessments for informed decision‐making in relation to soil management.     

Stabilization of dissolved organic matter by aluminium: a toxic effect or stabilization through precipitation?

Carbon mineralization in acidic forest soils can be retarded by large concentrations of aluminium (Al). However, it is still unclear whether Al reduces C mineralization by direct toxicity to microorganisms or by decreased bioavailability of organic matter (OM) because dissolved organic matter (DOM) is precipitated by Al. We conducted an incubation experiment (6 weeks) with two DOM solutions (40 mg C litre^?1) derived from two acidic forests and possessing large differences in composition. Aluminium was added to the solutions in realistic ranges for acidic soils (1.6–24 mg Al litre^?1) at pHs of 3.8 and 4.5, to achieve differences in Al speciation. We determined different Al species, including the potentially toxic Al³⁺, by Diffusive Gradients in Thin Films (DGT) to evaluate toxic effects on microorganisms. Precipitation of OM increased with larger amounts of added Al and higher pH, and we measured a larger fraction of dissolved ‘free’ Al at pH 3.8 than at pH 4.5. Organic matter degradation decreased significantly with Al addition, and we found more organic matter degraded at pH 3.8 than at pH 4.5 for the respective Al additions. Consequently, the observed reduction in OM degradation (i.e. stabilization) cannot be explained by toxic effects of ‘free’ Al. However, C stabilization correlated significantly with C precipitation. The pH did not influence C stabilization directly, but determined the amount of C being precipitated. Phosphorus was removed along with OM by precipitation, which possibly also affected C stabilization. We conclude that C stabilization upon Al addition did not result from toxic effects, but was caused by reduced bioavailability of OM after its precipitation. The reduction in OM degradation by 65% is of great relevance for the overall C stabilization in acidic forest soils. Increasing pH and decreasing Al concentrations upon recovery from acidic deposition should therefore not result in decreased stabilization of precipitated OM.     

A general method for measuring relative change in range size from biological atlas data

Conservationists need information on trends in the ranges of species in order to set priorities and to assign threat statuses using IUCN criteria. For many taxonomic groups in many countries, the best data available for this purpose come from atlases that map the distributions of species in grid cells. This paper presents a method of calculating an index of relative change in range size using atlas data for two periods. The method allows for the effect of variations in the geographical coverage and intensity of recorder effort. A weighted linear regression model is fitted to the relationship between counts of grid cells (as logit-transformed proportions) in the earlier and later periods. The standardised residual for each species provides an index of its relative change in range size. The method is illustrated by analyses of three British data sets. Previous methods of assessing change from atlas data are briefly reviewed and the advantages and limitations of the new method are discussed.     

Quantification of Organic Matter in Agricultural Soils from the Central Region of Paraná State,Brazil

The organic matter (OM) quantity in soils is of fundamental importance for agriculture. The indirect determination of the OM through the Total Organic Carbon (TOC) quantity is performed by most soil laboratories in Brazil using the Walkley–Black (WB) method. This procedure involves oxidation with potassium dichromate which is a cancerous reagent. The objective of this study is to optimize the parameters for OM determination by the gravimetric method and to estimate the van Bemmelen factor for the studied soil. The studied region is the second and third plateau of Parana State, Brazil, from which 50 agricultural soil samples were analyzed. The temperature and exposition time in muffle were determined after a thermal analysis. The optimized parameters for the gravimetric method were 3 h at 420°C in the muffle furnace. The results for WB and gravimetric methods presented a good correlation and the van Bemmelen factor for the studied soil was 4.37.     

Nuclear Magnetic Resonance,Infra-Red and Pyrolysis: Application of Spectroscopic Methodologies to Maturity Determination of Composts

Compost maturity or stability reflects the degree of decomposition of the organic matter (OM). Since stability of natural OM is a relative term, defining it is not a trivial challenge. In addition, it requires a series of chemical, physico-chemical and spectroscopic determinations. Among the methods applied, ¹³C-NMR and FTIR (or DRIFT) and pyrolysis have been shown to be of significance and therefore this review will be dedicated to studies focusing on the application of these methods to composting research. In fact, solid-state ¹³C-NMR spectroscopy has become the most important tool for examining the chemical structure of natural OM (NOM) and the chemical changes associated with OM decomposition. Changes can be measured on the bulk OM either fresh or composted, on humic substances (HS) extracted from the compost or on dissolved organic matter (DOM). Recently, 2D ¹H NMR has been employed to study properties of HS extracted from MSW compost. In general, changes measured on decomposing OM are more distinct in the following order of tested materials: DOM > Bulk OM > HS > Core HS. In conclusion, compost HS which are “young” relative to soil HS were shown to differ from the latter mostly in their high levels of aliphatic and polysaccharide components, which tend to decompose during composting. ¹³C-NMR is the most effective instrument applied to date to structural studies of NOM.     

How air-drying and rewetting modify soil organic matter characteristics: An assessment to improve data interpretation and inference

Air-drying and wetting of air-dried soil samples with water (i.e., rewetting) are widely used sample treatments in soil analyses. It is recognized that both air-drying and rewetting of soil samples affect the characteristics of organic matter (OM), but systematic evaluations are scarce. In this review, we synthesize what is known in the scientific literature concerning the types and magnitudes of effects resulting from air-drying and rewetting with respect to i) characteristics of aggregate-associated and water-extractable OM, ii) soil microbiota, and iii) decomposition of OM. Air-drying of soil samples results in the formation of new and/or stronger OM-mineral interactions as well as increased hydrophobicity and mineral surface acidity. The formation of new and enhancement of existing OM-mineral interactions may lead to an increase in perceived aggregate stability, potentially affecting estimates of amount and persistence of OM associated with soil aggregates. Compared to field moist samples, air-dried samples had 8–41% higher relative dry mass proportions in the 2–0.25 mm aggregate size fraction. Pronounced changes in the amount and composition of the water-extractable OM and soil microbiota are also detected during the course of air-drying and rewetting with the potential to affect the conclusions derived from OM decomposition experiments. Air-dried soil samples were found to have 2–10 times higher amounts of water extractable organic carbon and a decrease between 3% and 69% in the microbial biomass carbon (using the substrate-induced respiration technique) compared to field moist samples. The magnitude of air-drying and rewetting derived effects on sample characteristics appears to be site and soil type specific.     

Theoretical approaches to modelling the dynamics of soil organic matter

Two theoretical concepts of the formation of the soil organic matter (OM) system are considered as a methodological basis for the mathematical simulation of its dynamics. In the theory of physical protection of OM without the formation of humic substances with the physical fractionation methodolody, the main focus is on the functional parameters of the OM components separated by their mineralization rates. Here, two conceptual disadvantages are noted: (a) neglect of the specificity of OM transformation in organic soil horizons, where humified OM resistant to mineralization is formed, which cannot be explained by this theory; and (b) consideration of the soil microorganisms as a unified undifferentiated complex. In classical humification theory, a number of humification stages are considered with the respective communities of decomposer organisms that mineralize OM and transform biota products into humic substances. The silvicultural concept of humus forms was found to be effective and suitable for a wide range of natural conditions with the use of this theory for OM simulation. There is a general shortcoming to both approaches: protection theory has no parameters of recalcitrant OM formation from other fractions; in humification theory, the quantitative humification parameters under the effect of soil fauna have yet to be sufficiently substantiated. The values of the turnover time for active, intermediate, and recalcitrant OM are revised. The importance of theoretically substantiating the structural and functional organization of OM for its dynamic modelling is emphasized.     

无机肥料中有效磷含量的测定国际标准制定研究

为统一全球无机肥料中有效磷含量的测定方法,对国际上现有的3种主要检测方法进行了分析研究,比较了我国现行仲裁方法(GB/T 8573)、欧盟标准方法(EN 15959)以及传统国际标准方法(ISO 6598)的差异性,研究表明上述3种方法对测定结果没有显著差异.ISO/TC 134推荐了以GB/T 8573为基础,制订新...     

A fuzzy logic-classification of sediments based on data from in vitro biotests

Background, aim, and scope  Ecotoxicological risk assessment of sediments is usually based on a multitude of data obtained from tests with different endpoints. In the present study, a fuzzy logic-based model was developed in order to reduce the complexity of these data sets and to classify sediments on the basis of results from a battery of in vitro biotests. Materials and methods  The membership functions were adapted to fit the specific sensitivity and variability of each biotest. For this end, data sets were categorized into three toxicity levels using the box plot and empirical methods. The variability of each biotest was determined to calculate the range of the gradual membership. In addition, the biotests selected were ranked according to the biological organisation level in order to consider the ecological relevance of the endpoints measured by selected over- or underestimation of the toxicity levels. In the next step of the fuzzy logic model, a rule-base was implemented using if...and...then decisions to arrive at a system of five quality classes. Results  The results of the classification of sediments from the Rhine and Danube Rivers showed the highest correlation between the biotest results and the fuzzy logic alternative based on the empirical method (i.e. the classification of the data sets into toxicity levels). Discussion  Many different classification systems based on biological test systems are depending on respective data sets; therefore, they are difficult to compare with other locations. Furthermore, they don‘t consider the inherent variability of biotests and the ecological relevance of these test systems as well. In order to create a comprehensive risk assessment for sediments, mathematical models should be used which take uncertainties of biotest systems into account, since they are of particular importance for a reliable assessment. In the present investigation, the variability and ecological relevance of biotests were incorporated into a classification system based on fuzzy logic. Furthermore, since data from different sites and investigations were used to create membership functions of the fuzzy logic, this classification system has the potential to be independent of locations. Conclusions  In conclusion, the present fuzzy logic classification model provides an opportunity to integrate expert knowledge as well as acute and mechanism-specific effects for the classification of sediments for an ecotoxicological risk assessment. Recommendations and perspectives  In order to achieve a more comprehensive classification, further investigation is needed to incorporate results of chemical analyses and in situ parameters. Furthermore, more discussions are necessary with respect to the relative weight attributed to different ecological and chemical parameters in order to obtain a more precise assessment of sediments.     

Effects of nitrogen supply on water‐use efficiency of higher plants

The worldwide increase of food demand and reduced sweet‐water availability in some important food‐producing regions raised interest in more efficient water use, which has become one of the central research topics in agriculture. Improved irrigation management and reduced bare‐soil evaporation have highest priority to increase agronomic water‐use efficiency (WUE). Compared to these technical (irrigation) and basic (crop production) management options, effects of nutrient management on WUE were less frequently considered. Twenty‐nine publications on nitrogen (N) effects on biomass WUE of container‐grown plants are considered in this review. Most of them indicate positive N effects on WUE, and relevance of N effects on intrinsic WUE and unproductive water and carbon loss is discussed. A plot of 90 published data of percent decreases of WUE and dry mass under variable N supply is presented. Extrapolation of biomass WUE from leaf measurements of intrinsic WUE is critically reviewed. The positive correlation between WUE and dry‐mass formation suggests that physiological rather than stomatal effects are more important in order to explain positive N effects on WUE.     

A Chronic Plant Test for the Assessment of Contaminated Soils. Part 1: Method development (9 pp)

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PREAMBLE According to the German Federal Soil Protection Act (BBodSchG 1998), the habitat function of soils must be protected. Despite the fact that in the Federal Soil Protection Ordinance (BBodSchV 1999) it has not been established how this goal can be reached reliably, it is clear that such a biological function can only adequately be assessed using biological test methods. This is especially true when a soil is contaminated by a mixture of often unknown chemicals. In such a case the use of chemical analysis aiming at a small range of known substances is not sufficient and must therefore be supplemented by biological methods. For this reason, several standardised test methods are available (e.g. using earthworms, collembolans or plants; Römbke and Knacker 2003; ISO 2003). Since acute tests are usually not sensitive enough for the assessment of potentially contaminated soils (e.g. Hund-Rinke et al. 2002), chronic tests like the earthworm reproduction tests (ISO 1998) are recommended for this purpose.

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A chronic plant test for the determination of phytotoxicity was missing until quite recently. The term phytotoxicity is understood here as the capacity of a compound or a contaminated soil to cause temporary or long-lasting damage to plants (EPPO 1997). Therefore, the German Ministry for Education and Research sponsored a project (1997 – 1999) in which – based on existing standardised methods – such a chronic plant laboratory test was developed and partly validated (Kalsch and Römbke 2000). The new test can be used for the evaluation of single chemicals (see Part 1 of this mini-series) as well as for the assessment of contaminated or remediated soils (see Part 2 of this mini-series).

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ABSTRACT Background and Scope. As part of the efforts to improve the biological testing of contaminated soils, the German government sponsored the standardisation of a chronic plant bioassay. This new test is based on experiences with various acute plant tests (e.g. published by OECD or ISO) and existing North American Plant-Life-Cycle Bioassays. In this contribution the characteristic properties of the test are described.

Methods

The test can be performed either with Brassica rapa (turnip) or Avena sativa (oat). Its duration is 35 to 64 days with OECD artificial soil and a German standard field soil acting as controls. Water and nutrients are provided by an automatic wick irrigation system. Besides measuring biomass and shoot length, the number of pods, seeds and flowers are applied as chronic measurement endpoints. During the development of the test, TNT (2,4,6-trinitrotoluene) and Pyrene were used as model test substances.

Results

Pyrene did not affect B. rapa (turnip) in concentrations of up to 10.000 mg/kg soil (due to the often low sensitivity of A. sativa (oat) no further test with this substance was performed). Depending on the endpoint the results varied in the tests with TNT. With few exceptions, the NOEC (No Observed Effect Concentration) values were determined as 55.5 mg TNT/kg soil for B. rapa (turnip) and as 75 (unfertilised) and 150 (fertilised) mg TNT/kg soil for A. sativa (oat). The EC50-values varied between 96.3 and 207.2 mg TNT/kg soil for B. rapa (turnip) and 183.1 – 505.6 mg TNT/kg soil for A. sativa (oat), depending on the endpoint.

Outlook

The results of this work have been used to prepare a draft test guideline, which has recently been standardised by the International Organisation for Standardisation (ISO). Practical experiences with this test system are described in Part II of this mini series.     

有机无机肥长期定位试验土壤小麦季氨挥发损失及其影响因素研究

氨挥发是肥料氮素损失的重要途径之一,损失率因土壤类型、气候条件、肥料用量、施肥时间和方式等不同而存在很大差异。为了筛选提高氮肥利用率的肥料运筹方式,本文利用长期定位试验平台,采用间歇密闭通气法,研究了有机无机肥长期施用条件下小麦季土壤氨挥发损失及其影响因素。结果表明,不同肥料种类和配施强烈地影响着土壤氨挥发,在150kgN·hm^-2用量下小麦季氨挥发损失量以NK和有机肥处理为最高,分别达到17．89和15．70kgN·hm^-2,占氮肥用量的10．47％-11．93％,显著高于NPK、NP和有机无机肥配施（1／20M）处理。土壤氨挥发速率与气温呈显著正相关,基肥施用后灌水可以有效地降低氨挥发损失。NPK肥料平衡施用或者有机无机肥配施可以减少氨挥发损失。