This document specifies the crucial steps of a quantitative polymerase chain reaction (qPCR) method to measure the abundance of selected microbial gene sequences from soil DNA extract. The number of microbial gene sequences quantified by qPCR provides an estimation of the abundance of selected microbial groups in soil.

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This document specifies the crucial steps of a quantitative polymerase chain reaction (qPCR) method to measure the abundance of selected microbial gene sequences from soil DNA extract. The number of microbial gene sequences quantified by qPCR provides an estimation of the abundance of selected microbial groups in soil.

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This document deals with the assessment of human exposure to metals and metalloids from ingestion of soil. It specifies a simplified method for the prediction of the oral bioaccessibility of metals and metalloids from contaminated soils. The method is a chemical extraction using hydrochloric acid (HCl) and can be used at least for first-tier screening for prediction of the oral bioaccessibility of arsenic (As), cadmium (Cd) and lead (Pb) in soil samples. It produces extracts that are representative of the bioaccessible concentrations measured by using the validated unified bioaccessibility method (UBM) (ISO 17924). More specifically, from the extractable concentrations of metals and metalloids by HCl, the predicted bioaccessibility in both the gastric and gastrointestinal phases is calculated using the equations and the prediction intervals provided in this document, and which have been derived from simple linear regressions of data obtained during validation studies.[ REF Reference_ref_2 \r \h 2 08D0C9EA79F9BACE118C8200AA004BA90B0200000008000000100000005200650066006500720065006E00630065005F007200650066005F0032000000 ] NOTE The simplified method is applicable to prediction of the metals and metalloids bioaccessibility in soil with total concentrations range from 2 mg/kg to 2 600 mg/kg for As, from 0,2 mg/kg to 480 mg/kg for Cd, and from 4 mg/kg to 50 000 mg/kg for Pb.

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This document specifies a method for sampling, extracting and preserving macro-invertebrates from soils, including the litter zone.
The sampling and extraction methods in this document are applicable to almost all types of soil, with the exception of soils in extreme climatic conditions (hard, frozen or flooded soils) and matrices other than soil, e.g. tree trunks, plants or lichens.

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This document specifies a rapid method for assessing solid samples in an aerobic suspension, by determining the inhibition of dehydrogenase activity of Arthrobacter globiformis using the redox dye resazurin.
It is applicable for assessing the effect of water-soluble and solid matter bounded non-volatile contaminants in natural samples, such as soils and waste materials. Although not the main purpose, the contact test can additionally be used for testing the effect of chemicals, as described in the Annex C. The test yields a result within 6 h and can therefore be used for screening potentially contaminated test material.

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This document specifies a method for sampling, extracting and preserving collembolans and mites from field soils as a prerequisite for using these animals as bio-indicators (e.g. to assess the quality of a soil as a habitat for organisms).
The sampling and extraction methods of this document are applicable to almost all types of soils. Exceptions can be soils from extreme climatic conditions (hard, frozen or flooded soils) and other matrices than soil, e.g. tree trunks, plants or lichens.

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This document specifies test procedures for the determination of effects of contaminated soils or other contaminated samples on the emergence of lettuce seeds. This document is applicable to contaminated soils, soil materials, compost, sludge and chemical testing. It is also applicable to the measurement of effects of substances deliberately added to the soil and to the comparison of soils of known and unknown quality. This document is not applicable for volatile contaminants.

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This document specifies a method for sampling, extracting and preserving macro-invertebrates from soils, including the litter zone.
The sampling and extraction methods in this document are applicable to almost all types of soil, with the exception of soils in extreme climatic conditions (hard, frozen or flooded soils) and matrices other than soil, e.g. tree trunks, plants or lichens.

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This document specifies a method for sampling, extracting and preserving macro-invertebrates from soils, including the litter zone. The sampling and extraction methods in this document are applicable to almost all types of soil, with the exception of soils in extreme climatic conditions (hard, frozen or flooded soils) and matrices other than soil, e.g. tree trunks, plants or lichens.

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This document specifies a method for testing the bioaccessibility of substances from contaminated soil and soil-like materials. The method is not applicable for volatile contaminants. Furthermore, the method is only applicable if suitable analytical methods for extraction and detection of substances and/or elements from complex digestion assays are available. NOTE During the in vivo validation with minipigs, the PAHs naphthalene, acenaphthylene, acenaphthene and fluorene were not evaluated due to their volatility. However, the results of the overall recovery indicate if volatilisation has occurred during the test.

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This document specifies a rapid method for assessing solid samples in an aerobic suspension, by determining the inhibition of dehydrogenase activity of Arthrobacter globiformis using the redox dye resazurin.
It is applicable for assessing the effect of water-soluble and solid matter bounded non-volatile contaminants in natural samples, such as soils and waste materials. Although not the main purpose, the contact test can additionally be used for testing the effect of chemicals, as described in the Annex C. The test yields a result within 6 h and can therefore be used for screening potentially contaminated test material.

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This document specifies one of the methods for evaluating the habitat function of soils and determining effects of soil contaminants and substances on the reproduction of Enchytraeus sp. by dermal and alimentary uptake in a chronic test. It is applicable to soils and soil materials of unknown quality, for example, from contaminated sites, amended soils, soils after remediation, agricultural or other sites under concern and waste materials.
This document provides information on how to use this method for testing substances under temperate conditions.
The method is not applicable to substances, for which the air/soil partition coefficient is greater than 1, or to substances for which the vapour pressure exceeds 300 Pa at 25 °C.
NOTE      No provision is made in the test method for monitoring the persistence of the substance under test.

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This document specifies one of the methods for evaluating the habitat function of soils and determining effects of soil contaminants and substances on the reproduction of Folsomia candida Willem by dermal and alimentary uptake. This document also provides information on how to use this method for testing substances under temperate conditions.
The chronic test described is applicable to soils and soil materials of unknown quality, e.g. from contaminated sites, amended soils, soils after remediation, industrial, agricultural or other sites of concern and waste materials.
The method is not applicable to volatile substances, i.e. substances for which H (Henry's constant) or the air/water partition coefficient is greater than 1, or for which the vapour pressure exceeds 300 Pa at 25 °C.

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This document specifies a rapid method for assessing solid samples in an aerobic suspension, by determining the inhibition of dehydrogenase activity of Arthrobacter globiformis using the redox dye resazurin. It is applicable for assessing the effect of water-soluble and solid matter bounded non-volatile contaminants in natural samples, such as soils and waste materials. Although not the main purpose, the contact test can additionally be used for testing the effect of chemicals, as described in the Annex C. The test yields a result within 6 h and can therefore be used for screening potentially contaminated test material.

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This document specifies a method for sampling, extracting and preserving collembolans and mites from field soils as a prerequisite for using these animals as bio-indicators (e.g. to assess the quality of a soil as a habitat for organisms).
The sampling and extraction methods of this document are applicable to almost all types of soils. Exceptions can be soils from extreme climatic conditions (hard, frozen or flooded soils) and other matrices than soil, e.g. tree trunks, plants or lichens.

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This document specifies a method for sampling, extracting and preserving collembolans and mites from field soils as a prerequisite for using these animals as bio-indicators (e.g. to assess the quality of a soil as a habitat for organisms). The sampling and extraction methods of this document are applicable to almost all types of soils. Exceptions can be soils from extreme climatic conditions (hard, frozen or flooded soils) and other matrices than soil, e.g. tree trunks, plants or lichens.

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This document specifies one of the methods for evaluating the habitat function of soils and determining effects of soil contaminants and substances on the reproduction of Folsomia candida Willem by dermal and alimentary uptake. This document also provides information on how to use this method for testing substances under temperate conditions.
The chronic test described is applicable to soils and soil materials of unknown quality, e.g. from contaminated sites, amended soils, soils after remediation, industrial, agricultural or other sites of concern and waste materials.
The method is not applicable to volatile substances, i.e. substances for which H (Henry's constant) or the air/water partition coefficient is greater than 1, or for which the vapour pressure exceeds 300 Pa at 25 °C.

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This document specifies one of the methods for evaluating the habitat function of soils and determining effects of soil contaminants and substances on the reproduction of Folsomia candida Willem by dermal and alimentary uptake. This document also provides information on how to use this method for testing substances under temperate conditions. The chronic test described is applicable to soils and soil materials of unknown quality, e.g. from contaminated sites, amended soils, soils after remediation, industrial, agricultural or other sites of concern and waste materials. The method is not applicable to volatile substances, i.e. substances for which H (Henry's constant) or the air/water partition coefficient is greater than 1, or for which the vapour pressure exceeds 300 Pa at 25 °C.

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This document specifies one of the methods for evaluating the habitat function of soils and determining effects of soil contaminants and substances on the reproduction of Enchytraeus sp. by dermal and alimentary uptake in a chronic test. It is applicable to soils and soil materials of unknown quality, for example, from contaminated sites, amended soils, soils after remediation, agricultural or other sites under concern and waste materials.
This document provides information on how to use this method for testing substances under temperate conditions.
The method is not applicable to substances, for which the air/soil partition coefficient is greater than 1, or to substances for which the vapour pressure exceeds 300 Pa at 25 °C.
NOTE      No provision is made in the test method for monitoring the persistence of the substance under test.

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This document specifies one of the methods for evaluating the habitat function of soils and determining the effects of soil contaminants and chemicals on the reproduction of Eisenia fetida/Eisenia andrei by dermal and alimentary uptake. This chronic test is applicable to soils and soil materials of unknown quality, e.g. from contaminated sites, amended soils, soils after remediation, agricultural or other sites concerned, and waste materials.
This method is designed mainly for determining the effects of soil contaminants and chemicals on the reproduction of Eisenia fetida/Eisenia andrei. Technical information is also provided on how to use Eisenia fetida/andrei for testing chemicals under tropical conditions (see Annex A). Finally, this method also includes technical information on how to use it with other environmentally relevant earthworm species: e.g. Dendrodrilus rubidus and Aporrectodea caliginosa (see Annexes B and C).
This method does not apply to substances for which the air/soil partition coefficient is greater than one, or to substances with vapour pressure exceeding 300 Pa, at 25 °C. This method does not take into account the persistence of the substance during the test.

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This document specifies one of the methods for evaluating the habitat function of soils and determining effects of soil contaminants and substances on the reproduction of Enchytraeus sp. by dermal and alimentary uptake in a chronic test. It is applicable to soils and soil materials of unknown quality, for example, from contaminated sites, amended soils, soils after remediation, agricultural or other sites under concern and waste materials. This document provides information on how to use this method for testing substances under temperate conditions. The method is not applicable to substances, for which the air/soil partition coefficient is greater than 1, or to substances for which the vapour pressure exceeds 300 Pa at 25 °C. NOTE No provision is made in the test method for monitoring the persistence of the substance under test.

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SIGNIFICANCE AND USE
5.1 Spectral analysis of soils for agricultural use is being used worldwide to obtain rapid data on soil nutrients. for the purpose of agricultural management including fertilizer application and other amendments such as pH adjustment, organic supplements, etc. Satellite, aerial, and ground-based sampling methods are being used. This test method applies to ground-based, terrestrial field applications where samples are taken from the ground, generally in the root zone. Use of these rapid remote sensing techniques allow for more detailed and economic data acquisition than older cumbersome sampling and wet chemistry testing methods used in the past by soil scientists for soil nutrient evaluations.  
5.2 This test method describes procedures for sampling and testing of field soils using diffuse reflectance spectrometry using handheld portable spectrometers measuring spectra in visible and near infrared (vis-NR) using dried sieved or wet samples. There is a worldwide effort to collect spectral databases of soils. The procedures specified here follow procedures as outlined in the United Nations Food and Agricultural Organization (FAO) primer on Vis-NIR and MIR spectroscopy of soils (1)3. Other organizations such as IEEE are actively working on additional guidance documents that will be incorporated in future revisions of this test method.  
5.2.1 This standard describes the procedures (Section 12) for using hyperspectral sensor data to measure moisture content as a percentage, pH, Organic Matter (OM) as a percentage, Cation Exchange Capacity (CEC) measured in 10 cmol c /kg could hold 10 cmol of Na + cations (with 1 unit of charge per cation) per kilogram of soil, but only 5 cmol Ca 2+ (2 units of charge per cation), as well as micro and macro nutrients in soils measured in PPM (parts per million)or a percentage, including, but not limited to nitrogen, phosphorous, potassium, boron, zinc, iron, sulfur, calcium, magnesium, and manganese.  
5.2.2 Research has shown that t...
SCOPE
1.1 This test method describes procedures for sampling and testing of soils obtained from ground-based samples using diffuse reflectance spectrometry using handheld portable spectrometers measuring spectra in visible and near infrared (vis-NR) and mid-infrared (MIR) range. The sensor can measure moisture content, PH, organic matter, Cation Exchange Capacity (CEC) as well as macro and micro elemental nutrients in parts per million (PPM) or percentage, including but not limited to nitrogen, phosphorous, potassium, zinc, iron, boron, sulfur, calcium, magnesium, and manganese.  
1.2 There are two methods that can be used to perform the test.  
1.2.1 Method A—The analysis is performed in the laboratory on the sample after the sample has been oven dried and sieved.  
1.2.2 Method B—The analysis is performed in the field on a moist sample after homogenization. After post-processing of multiple reflectance site data using methods A and B, the moisture content can be measured, and the spectral signature is normalized for moisture content.  
1.3 The limitation of this method is that the results of an individual test for elemental analysis would not be the same as exacting reference values from traditional wet chemical lab analysis used by soil scientists. Results of wet chemistry tests or tests from soil science libraries may be used to calibrate a specific site model comprised of many individual tests. Spectral data for organics has shown to be as accurate as conventional methods such as Test Methods D2974.  
1.4 For soil nutrient analysis the sample is not finely ground as in typical qualitative spectral analysis as outlined in standard Practice E1252. The spectrometer is checked periodically during testing using procedures in accordance with Guide E1866 performance testing.  
1.5 Moisture content is a preferred term in agricultural applications. For this standard, gravimetric water content may be measured in accordance wi...

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This document specifies a method for sampling and handling free-living nematodes from terrestrial field soils as a prerequisite for using them as bio-indicators (e.g. to assess the quality of a soil as a habitat for organisms).
This document applies to all terrestrial biotopes in which nematodes occur. The sampling design of field studies in general is specified in ISO 18400-101.
This document is not applicable to aquatic nematodes because of differences in the sample matrix (e.g. water column). Methods for some other soil organism groups such as earthworms, collembolans enchytraeids or macro-invertebrates are covered in ISO 23611-1, ISO 23611-2, ISO 23611-3 and ISO 23611-5.
This document does not cover the pedological characterization of the site which is highly recommendable when sampling soil invertebrates. ISO 10390, ISO 10694, ISO 11272, ISO 11274, ISO 11277, ISO 11461 and ISO 11465 include suitable procedures for measuring pH, particle size distribution, C/N ratio, organic carbon content and water-holding capacity.

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This document specifies one of the methods for evaluating the habitat function of soils and determining the effects of soil contaminants and chemicals on the reproduction of Eisenia fetida/Eisenia andrei by dermal and alimentary uptake. This chronic test is applicable to soils and soil materials of unknown quality, e.g. from contaminated sites, amended soils, soils after remediation, agricultural or other sites concerned, and waste materials.
This method is designed mainly for determining the effects of soil contaminants and chemicals on the reproduction of Eisenia fetida/Eisenia andrei. Technical information is also provided on how to use Eisenia fetida/andrei for testing chemicals under tropical conditions (see Annex A). Finally, this method also includes technical information on how to use it with other environmentally relevant earthworm species: e.g. Dendrodrilus rubidus and Aporrectodea caliginosa (see Annexes B and C).
This method does not apply to substances for which the air/soil partition coefficient is greater than one, or to substances with vapour pressure exceeding 300 Pa, at 25 °C. This method does not take into account the persistence of the substance during the test.

  • Standard
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This document specifies one of the methods for evaluating the habitat function of soils and determining the effects of soil contaminants and chemicals on the reproduction of Eisenia fetida/Eisenia andrei by dermal and alimentary uptake. This chronic test is applicable to soils and soil materials of unknown quality, e.g. from contaminated sites, amended soils, soils after remediation, agricultural or other sites concerned, and waste materials. This method is designed mainly for determining the effects of soil contaminants and chemicals on the reproduction of Eisenia fetida/Eisenia andrei. Technical information is also provided on how to use Eisenia fetida/andrei for testing chemicals under tropical conditions (see Annex A). Finally, this method also includes technical information on how to use it with other environmentally relevant earthworm species: e.g. Dendrodrilus rubidus and Aporrectodea caliginosa (see Annexes B and C). This method does not apply to substances for which the air/soil partition coefficient is greater than one, or to substances with vapour pressure exceeding 300 Pa, at 25 °C. This method does not take into account the persistence of the substance during the test.

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SIGNIFICANCE AND USE
6.1 Contaminated sediments may affect natural populations of aquatic organisms adversely. Sediment-dwelling organisms may be exposed directly to contaminants by the ingestion of sediments and by the uptake of sediment-associated contaminants from interstitial and overlying water. Contaminated sediments may affect water column species directly by serving as a source of contaminants to overlying waters or a sink for contaminants from overlying waters. Organisms may also be affected when contaminated sediments are suspended in the water column by natural or human activities. Water column species and nonaquatic species may also be affected indirectly by contaminated sediments by the transfer of contaminants through ecosystems (7, 8).  
6.2 The procedures described in this guide may be used and adapted for incorporation in basic and applied research to determine the ecological effects of contaminated sediments. These same methods may also be used in the development and implementation of monitoring and regulatory programs designed to prevent and manage sediment contamination.  
6.3 Sediment tests with aquatic organisms can be used to quantify the acute and chronic toxicity and the bioavailability of new and presently used materials. Sediment toxicity may also result from environmental processes such as ammonia generation, pH shifts, or dissolved oxygen fluctuation. In many cases, consideration of the adverse effects of sediment-associated contaminants is only one part of a complete hazard assessment of manufactured compounds that are applied directly to the environment (for example, pesticides) and those released (for example, through wastewater effluents) as by-products from the manufacturing process or from municipalities (7).  
6.4 Sediment tests can be used to develop exposure-response relationships for individual toxicants by spiking clean sediments with varying concentrations of a test chemical and determining the concentration that elicits the target response in...
SCOPE
1.1 As the contamination of freshwater and saltwater ecosystems continues to be reduced through the implementation of regulations governing both point and non-point source discharges, there is a growing emphasis and concern regarding historical inputs and their influence on water and sediment quality. Many locations in urban areas exhibit significant sediment contamination, which poses a continual and long-term threat to the functional condition of benthic communities and other species inhabiting these areas (1).2 Benthic communities are an important component of many ecosystems and alterations of these communities may affect water-column and nonaquatic species.  
1.2 Biological tests with sediments are an efficient means for evaluating sediment contamination because they provide information complementary to chemical characterizations and ecological surveys (2). Acute sediment toxicity tests can be used as screening tools in the early phase of an assessment hierarchy that ultimately could include chemical measurements or bioaccumulation and chronic toxicity tests. Sediment tests have been applied in both saltwater and freshwater environments (2-6). Sediment tests have been used for dredge material permitting, site ranking for remediation, recovery studies following management actions, and trend monitoring. A particularly important application is for establishing contaminant-specific effects and the processes controlling contaminant bioavailability(7).  
1.3 This guide is arranged as follows:    
Section  
Referenced Documents  
2  
Terminology  
3  
Application  
4  
Summary of Guide  
5  
Significance and Use  
6  
Hazards  
7  
Sediment Test Types  
8  
Biological Responses  
9  
Test Organisms  
10  
Experimental Design Considerations  
11  
Data Interpretation  
12  
Keywords  
13  
1.4 The values stated in SI units are to be regarded as the standard. The values g...

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This document specifies a test procedure for the evaluation of the habitat function of soils by determining effects of soil contaminants and substances on organic matter decomposition. This test is applicable to natural soils and soil materials of unknown quality (e.g. contaminated sites, amended soils, soils after remediation, agricultural or other sites under concern). This document also specifies how to use this method for testing substances under temperate conditions. This document is not applicable to substances for which the air/soil partition coefficient is greater than 1. It is not applicable to substances with vapour pressure exceeding 300 Pa at 25 °C. NOTE The stability of the test substance cannot be ensured over the test period. No provision is made in the test method for monitoring the persistence of the substance under test.

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This document aims to assist in designing and organizing trials for validation of biotests. The validation activities during the different steps of the standardization process are described. This document comprises the overall data evaluation and subsequent validation study conclusion. This document is intended for the validation of biotests which can differ in their experimental design and endpoints. It is possible that some of the requirements of this document are not applicable to all test methods.

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This document specifies a method for sampling and handling free-living nematodes from terrestrial field soils as a prerequisite for using them as bio-indicators (e.g. to assess the quality of a soil as a habitat for organisms).
This document applies to all terrestrial biotopes in which nematodes occur. The sampling design of field studies in general is specified in ISO 18400-101.
This document is not applicable to aquatic nematodes because of differences in the sample matrix (e.g. water column). Methods for some other soil organism groups such as earthworms, collembolans enchytraeids or macro-invertebrates are covered in ISO 23611-1, ISO 23611-2, ISO 23611-3 and ISO 23611-5.
This document does not cover the pedological characterization of the site which is highly recommendable when sampling soil invertebrates. ISO 10390, ISO 10694, ISO 11272, ISO 11274, ISO 11277, ISO 11461 and ISO 11465 include suitable procedures for measuring pH, particle size distribution, C/N ratio, organic carbon content and water-holding capacity.

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SIGNIFICANCE AND USE
5.1 Soil toxicity tests provide information concerning the toxicity and bioavailability of chemicals associated with soils to terrestrial organisms. As important members of the soil fauna, nematodes have a number of characteristics that make them appropriate organisms for use in the assessment of potentially hazardous soils. Bacterial-feeding nematodes such as C. elegans feed on soil microbes and contribute to the breakdown of organic matter. They are also of extreme importance in the cycling and degradation of key nutrients in soil ecosystems (9). Soil nematodes also serve as a source of prey and nutrients for fauna and microflora such as soil nematophagous fungi (10). A major change in the abundance of soil invertebrates such as nematodes, either as a food source or as organisms functioning properly in trophic energy transfer and nutrient cycling, could have serious adverse ecological effects on the entire terrestrial system.  
5.2 Results from soil tests might be an important consideration when assessing the hazards of materials to terrestrial organisms.  
5.3 The soil test might be used to determine the temporal or spatial distribution of soil toxicity. Test methods can be used to detect horizontal and vertical gradients in toxicity.  
5.4 Results of soil tests could be used to compare the sensitivities of different species.  
5.5 An understanding of the effect of these parameters on toxicity may be gained by varying soil characteristics such as pH, clay content, and organic material.  
5.6 Results of soil tests may be useful in helping to predict the effects likely to occur with terrestrial organisms in field situations.  
5.6.1 Field surveys can be designed to provide either a qualitative or quantitative evaluation of biological effects within a site or among sites.  
5.6.2 Soil surveys evaluating biological effects are usually part of more comprehensive analyses of biological, chemical, geological, and hydrographic conditions. Statistical correlation c...
SCOPE
1.1 This guide covers procedures for obtaining laboratory data to evaluate the adverse effects of chemicals associated with soil to nematodes from soil toxicity tests. This standard is based on a modification to Guide E1676. The methods are designed to assess lethal or sublethal toxic effects on nematodes in short-term tests in terrestrial systems. Soils to be tested may be (1) references soils or potentially toxic soil sites; (2) artificial, reference, or site soils spiked with compounds; (3) site soils diluted with reference soils; or (4) site or reference soils diluted with artificial soil. Test procedures are described for the species Caenorhabditis elegans (see Annex A1). Methods described in this guide may also be useful for conducting soil toxicity tests with other terrestrial species, although modifications may be necessary.  
1.2 Summary of Previous Studies—Initial soil toxicity testing using the free-living, bacterivorous soil nematode Caenorhabditis elegans was developed by Donkin and Dusenbery (1).2 Following the development of an effective method of recovery of C. elegans  from test soils, the organism was used to identify factors that affect the toxicity of zinc, cadmium, copper, and lead (2) . Freeman et al. further refined the nematode bioassay by decreasing the quantity of soil and spiking solution volumes, determining test acceptability criteria, and developing control charts to assess worm health using copper as a reference toxicant (3). More recently, the toxicological effects of nitrate and chloride metallic salts in two natural soils were compared (4) . LC50 values for C. elegans exposed for 24-h to nitrate salts of cadmium, copper, zinc, lead and nickel in an artificial soil (see Annex A2) were found to be similar to LC50 values for the earthworm, Eisenia fetida  (5). Increasing the exposure time to 48-h resulted in much lower LC50 values (6). However, longer exposure times necessitate the additio...

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This document specifies a method for sampling and handling free-living nematodes from terrestrial field soils as a prerequisite for using them as bio-indicators (e.g. to assess the quality of a soil as a habitat for organisms). This document applies to all terrestrial biotopes in which nematodes occur. The sampling design of field studies in general is specified in ISO 18400-101. This document is not applicable to aquatic nematodes because of differences in the sample matrix (e.g. water column). Methods for some other soil organism groups such as earthworms, collembolans enchytraeids or macro-invertebrates are covered in ISO 23611-1, ISO 23611-2, ISO 23611-3 and ISO 23611-5. This document does not cover the pedological characterization of the site which is highly recommendable when sampling soil invertebrates. ISO 10390, ISO 10694, ISO 11272, ISO 11274, ISO 11277, ISO 11461 and ISO 11465 include suitable procedures for measuring pH, particle size distribution, C/N ratio, organic carbon content and water-holding capacity.

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This document describes a method to assess the bioaccumulation of chemicals in snails, i.e. concentrations of metal(loid)s (ME) or organic compounds [e.g. polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs)] accumulated in their tissues.
This document presents how to prepare snails for caging in situ for 28 days, the in situ test design and then how to collect and prepare the snails until conservation and further analysis. If a kinetic study of accumulation is necessary, sampling of snails at different time-points during exposure is possible as well [13],[19],[22].
This document excludes analytical methods. Preparation (extraction and mineralization) of the samples and quantification of chemicals are not in the scope of the present document.
The method is applicable for soils under different uses (agricultural, industrial, residential, forests, before and after remediation, on potentially contaminated sites, etc.) and waste materials [8],[10], preferably with vegetation and/or humus cover.
The method is applicable subject to certain limits of temperature (frost-free period, i.e. mainly from April to October in temperate region).
Optionally (see Annex I), the method can be used in the laboratory to evaluate the accumulation of contaminants [and optionally, the sum of excess of transfer (SET) index for ME, PAH, PCB] of snails exposed only to soil.

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This document describes a method to assess the bioaccumulation of chemicals in snails, i.e. concentrations of metal(loid)s (ME) or organic compounds [e.g. polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs)] accumulated in their tissues.
This document presents how to prepare snails for caging in situ for 28 days, the in situ test design and then how to collect and prepare the snails until conservation and further analysis. If a kinetic study of accumulation is necessary, sampling of snails at different time-points during exposure is possible as well [13],[19],[22].
This document excludes analytical methods. Preparation (extraction and mineralization) of the samples and quantification of chemicals are not in the scope of the present document.
The method is applicable for soils under different uses (agricultural, industrial, residential, forests, before and after remediation, on potentially contaminated sites, etc.) and waste materials [8],[10], preferably with vegetation and/or humus cover.
The method is applicable subject to certain limits of temperature (frost-free period, i.e. mainly from April to October in temperate region).
Optionally (see Annex I), the method can be used in the laboratory to evaluate the accumulation of contaminants [and optionally, the sum of excess of transfer (SET) index for ME, PAH, PCB] of snails exposed only to soil.

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This document describes a method to assess the bioaccumulation of chemicals in snails, i.e. concentrations of metal(loid)s (ME) or organic compounds [e.g. polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs)] accumulated in their tissues. This document presents how to prepare snails for caging in situ for 28 days, the in situ test design and then how to collect and prepare the snails until conservation and further analysis. If a kinetic study of accumulation is necessary, sampling of snails at different time-points during exposure is possible as well [13],[19],[22]. This document excludes analytical methods. Preparation (extraction and mineralization) of the samples and quantification of chemicals are not in the scope of the present document. The method is applicable for soils under different uses (agricultural, industrial, residential, forests, before and after remediation, on potentially contaminated sites, etc.) and waste materials [8],[10], preferably with vegetation and/or humus cover. The method is applicable subject to certain limits of temperature (frost-free period, i.e. mainly from April to October in temperate region). Optionally (see Annex I), the method can be used in the laboratory to evaluate the accumulation of contaminants [and optionally, the sum of excess of transfer (SET) index for ME, PAH, PCB] of snails exposed only to soil.

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This document specifies one of the methods for evaluating the habitat function of soils and determining effects of soil contaminants and individual chemical substances on the reproduction of the oribatid mite Oppia nitens by dermal and alimentary uptake. This chronic (28-day) test is applicable to soils and soil materials of unknown quality (e.g., contaminated sites, amended soils, soils after remediation, agricultural or other sites under concern and waste materials). This method is not intended to replace the earthworm or Collembola tests since it represents another taxonomic group (= mites; i.e., arachnids), nor the predatory mite test since this species represents a different trophic level and ecological niche.
Effects of substances are assessed using standard soil, preferably a defined artificial soil substrate. For contaminated soils, the effects are determined in the test soil and in a control soil. According to the objective of the study, the control and dilution substrate (dilution series of contaminated soil) should be either an uncontaminated soil with similar properties to the soil sample to be tested (reference soil) or a standard soil (e.g., artificial soil).
Information is provided on how to use this method for testing substances under temperate conditions.
This document is not applicable to substances for which the air/soil partition coefficient is greater than 1, or to substances with vapour pressure exceeding 300 Pa at 25 °C.
NOTE    The stability of the test substance cannot be assured over the test period. No provision is made in the test method for monitoring the persistence of the substance under test.

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This document specifies a simple method for the extraction of only phospholipid fatty acids (PLFA) from soils.

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This document specifies a method for determining the toxicity of environmental samples on growth, fertility and reproduction of Caenorhabditis elegans. The method applies to contaminated whole freshwater sediment (maximum salinity 5 g/l), soil and waste, as well as to pore water, elutriates and aqueous extracts that were obtained from contaminated sediment, soil and waste.

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This document specifies a simple method for the extraction of only phospholipid fatty acids (PLFA) from soils.

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This document specifies a simple method for the extraction of only phospholipid fatty acids (PLFA) from soils.

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This document specifies one of the methods for evaluating the habitat function of soils and determining effects of soil contaminants and individual chemical substances on the reproduction of the oribatid mite Oppia nitens by dermal and alimentary uptake. This chronic (28-day) test is applicable to soils and soil materials of unknown quality (e.g., contaminated sites, amended soils, soils after remediation, agricultural or other sites under concern and waste materials). This method is not intended to replace the earthworm or Collembola tests since it represents another taxonomic group (= mites; i.e., arachnids), nor the predatory mite test since this species represents a different trophic level and ecological niche.
Effects of substances are assessed using standard soil, preferably a defined artificial soil substrate. For contaminated soils, the effects are determined in the test soil and in a control soil. According to the objective of the study, the control and dilution substrate (dilution series of contaminated soil) should be either an uncontaminated soil with similar properties to the soil sample to be tested (reference soil) or a standard soil (e.g., artificial soil).
Information is provided on how to use this method for testing substances under temperate conditions.
This document is not applicable to substances for which the air/soil partition coefficient is greater than 1, or to substances with vapour pressure exceeding 300 Pa at 25 °C.
NOTE    The stability of the test substance cannot be assured over the test period. No provision is made in the test method for monitoring the persistence of the substance under test.

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The present document specifies a method for direct extraction of DNA from soil samples to analyse the abundance and composition of microbial communities by various techniques of molecular biology including real-time quantitative PCR (qPCR). This method is mainly dedicated to agricultural and forest soils. This method can possibly not be suitable for soils rich in organic matter (e.g. peat soils) or soils heavily polluted with organic pollutants or heavy metals.
The direct extraction of DNA from soil samples provides unique insight into the α- and β-diversity of microbial communities. Next-generation sequencing of amplicons obtained by PCR (polymerase chain reaction) amplification of soil DNA constitutes a promising domain which will in the near future contribute to the development of routine tools to monitor microbial communities in soil environments.

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SIGNIFICANCE AND USE
5.1 Soil toxicity tests provide information concerning the toxicity and bioavailability of chemicals associated with soils to terrestrial organisms. As important members of the soil fauna, lumbricid earthworms and enchytraeid potworms have a number of characteristics that make them appropriate organisms for use in the assessment of potentially hazardous soils. Earthworms may ingest large quantities of soil, have a close relationship with other soil biomasses (for example, invertebrates, roots, humus, litter, and microorganisms), constitute up to 92 % of the invertebrate biomass of soil, and are important in recycling nutrients (1, 2).4 Enchytraeids contribute up to 5.2 % of soil respiration, constitute the second-highest biomass in many soils (the highest in acid soils in which earthworms are lacking) and effect considerably nutrient cycling and community metabolism (3-5). Earthworms and potworms accumulate and are affected by a variety of organic and inorganic compounds (2-10, 11-14). In addition, earthworms and potworms are important in terrestrial food webs, constituting a food source for a very wide variety of organisms, including birds, mammals, reptiles, amphibians, fish, insects, nematodes, and centipedes  (15, 16, 3). A major change in the abundance of soil invertebrates such as lumbricids or enchytraeids, either as a food source or as organisms functioning properly in trophic energy transfer and nutrient cycling, could have serious adverse ecological effects on the entire terrestrial system.  
5.2 A number of species of lumbricids and enchytraeid worms have been used in field and laboratory investigations in the United States and Europe. Although the sensitivity of various lumbricid species to specific chemicals may vary, from their study of four species of earthworms (including E. fetida) exposed to ten organic compounds representing six classes of chemicals, Neuhauser, et al  (7)  suggest that the selection of earthworm test species does not affect the a...
SCOPE
1.1 This guide covers procedures for obtaining laboratory data to evaluate the adverse effects of contaminants (for example, chemicals or biomolecules) associated with soil to earthworms (Family Lumbricidae) and potworms (Family Enchytraeidae) from soil toxicity or bioaccumulation tests. The methods are designed to assess lethal or sublethal toxic effects on earthworms or bioaccumulation of contaminants in short-term tests (7 to 28 days) or on potworms in short to long-term tests (14 to 42 days) in terrestrial systems. Soils to be tested may be (1) reference soils or potentially toxic site soils; (2) artificial, reference, or site soils spiked with compounds; (3) site soils diluted with reference soils; or (4) site or reference soils diluted with artificial soil. Test procedures are described for the species Eisenia fetida (see Annex A1) and for the species Enchytraeus albidus (see Annex A4). Methods described in this guide may also be useful for conducting soil toxicity tests with other lumbricid and enchytraeid terrestrial species, although modifications may be necessary.  
1.2 Modification of these procedures might be justified by special needs. The results of tests conducted using atypical procedures may not be comparable to results using this guide. Comparison of results obtained using modified and unmodified versions of these procedures might provide useful information concerning new concepts and procedures for conducting soil toxicity and bioaccumulation tests with terrestrial worms.  
1.3 The results from field-collected soils used in toxicity tests to determine a spatial or temporal distribution of soil toxicity may be reported in terms of the biological effects on survival or sublethal endpoints (see Section 14). These procedures can be used with appropriate modifications to conduct soil toxicity tests when factors such as temperature, pH, and soil characteristics (for example, particle size, organic matter c...

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1 Scope
This International Standard specifies a test method for determining the activity of active aerobic, heterotrophic microbial biomass in soils. This method is applicable to the monitoring of soil quality and to the evaluation of the ecotoxic potential of soils and soil materials. It is also applicable for soils sampled along contamination gradients in the field and to soils that are contaminated experimentally in the field or in the laboratory.

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ISO 17512-1:2008 specifies a rapid screening method for evaluating the habitat function of soils and the influence of contaminants and chemicals on earthworm behaviour.
The sublethal test is a rapid method that reflects the bioavailability of contaminant mixtures in natural soils and substances spiked into soils to Eisenia fetida and Eisenia andrei. The avoidance behaviour of the worms is the measurement endpoint of the test. This test is not intended to replace the earthworm reproduction test.
Two different designs (a two section unit and a six section unit) have been developed and successfully applied. Both designs are applicable to either single-concentration (e. g. for assessing the quality of a field soil) or multi-concentration (e. g. for assessing the toxicity of a spiked chemical) tests. In both cases, the earthworms are allowed to make the initial choice on which compartment, control and a treatment [in the two section test vessel between right and left side; in the six section test vessel between the (3 + 3) alternating compartments], to enter.

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The purpose of ISO 29200:2013 is to describe a method for assessing genotoxic effects (chromosome breakage or dysfunction of the mitotic spindle) of soils or soil materials on the secondary roots of a higher plant: Vicia faba (broad bean). This method allows the assessment of genotoxicity (toxicity for genetic material) of soils and soil materials like compost, sludge, waste, fertilizing matters, etc. Two ways of exposure can be considered: a direct exposure of plants to the soil (or soil material) which is relevant for the real genotoxic potential and an exposure of plants to the water extract of the soil (or soil material). This last way of exposure to a leachate or an eluate allows the detection of the mutagens which are not adsorbed to soils and which may be transferred to aquatic compartments. Moreover, this test may be used to evaluate genotoxic effects of chemical substances and to waters, effluents, etc.

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ISO 17512-2:2011 specifies a rapid screening method for evaluating the habitat function of soils based on the avoidance behaviour of springtails.
The test is a rapid method that reflects the bioavailability of contaminants in natural soils and substances spiked into soils to Folsomia candida. In both cases, it is possible to establish a dose-response-relationship. The avoidance behaviour of the springtails is the measurement endpoint of the test. This test is not intended to replace the Collembola reproduction test.

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