Some standard content:
ICS13.020
National Standard of the People's Republic of China
GB/T 27855-2011
Chemicals
Soil microorganisms
Carbon transformation test
ChemicalsSoil microorganisms--Carbon transformation test2011-12-30Issued
General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of ChinaStandardization Administration of the People's Republic of China
Implementation on August 1, 2012
This standard was drafted in accordance with the regulations given in GB/T1.1-2009. GB/T27855-2011
This standard has the same technical content as the Organization for Economic Cooperation and Development (OECD) Guidelines for Testing Chemicals 217 & Soil Microorganisms Carbon Transformation Test 3 (English version).
This standard has been revised as follows:
The original text of "INTRODUCTION\ and \INITIALCONSIDERATIONS\" are combined as the introduction of this standard; the scope is increased:
The content of \DEFINITIONS\ in the original text is used as the "Terms and Definitions" in the standard - The measurement units are uniformly changed to the legal measurement units of my country; This standard is proposed and managed by the National Technical Committee for Standardization of Hazardous Chemicals Management (SAC/TC251). The drafting units of this standard are: Hubei Entry-Exit Inspection and Quarantine Bureau, Guangdong Microbiological Analysis and Testing Center, Chemical Registration Center of the Ministry of Environmental Protection, and China Institute of Inspection and Quarantine. The main drafters of this standard are: Cui Hairong, Guo Jian, Zhao Hui, Cao Hedgehog, Liu Chunxin, Lian Huiming, Jian Yan, Yang Shunfeng, Ye Cheng1
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GB/T27B55—2011
This test method is designed to study the long-term potential effects of a single chemical on soil microbial carbon conversion activity. This test method is based on the test method recommended by the European and Mediterranean Plant Protection Organization1, and also refers to the test methods of the German Federal Institute for Biological Research, the US Environmental Protection Agency:*1, the Society of Environmental Toxicology and Chemistry and the International Organization for Standardization1. In 1995, the Soil/Sediment Working Group of the Organization for Economic Cooperation and Development held a meeting in Belgica, Italy-\1, and formally determined the types and quantities of soils used in this test. For the collection, processing, storage, etc. of soil samples, please refer to the guidance documents of IS) and the Belgica Working Group The impact of the chemical on soil microbial activity is measured when evaluating the phytochemical properties of a test substance, for example, when data are needed on the adverse effects of crop protection agents on soil microflora, or when the effects of exposure of soil microorganisms to chemicals other than agrochemicals are to be shown. Carbon conversion tests are designed to measure the effects of such chemicals on soil microbial flora. If the test substance is an agrochemical (e.g., pesticide, fertilizer, forestry chemical), both ammonia conversion and carbon conversion tests should be performed. If the test substance is not an agrochemical, only nitrogen conversion tests should be performed. However, if during the test, the nitrogen conversion ECs are found to be within the range of commercial nitrification inhibitors (e.g., 2-chloro-6-triazinemethylpyridine), carbon conversion tests should be performed to obtain further information. Soils consist of a complex, heterogeneous complex of biotic and abiotic organisms. Microbial organisms play a vital role in the decomposition and transformation of organic matter in fertile soils, and the combined action of different biological species can have a significant impact on various aspects of soil fertility. Any change in these biochemical processes has the potential to interfere with the nitrogen cycle in the long term, thereby altering soil fertility. Carbon and nitrogen transformations occur in all fertile soils. Although the contribution of microbial communities to these processes varies with soil type, the pathways of transformation are essentially the same.
This test method is used to detect the long-term adverse effects of a substance on carbon transformation processes on the soil surface under aerobic conditions: the test is very sensitive to changes in the number and activity of carbon-transferring microbial communities, which are affected by chemical and carbon stresses. Sand fills were used because of their low organic matter content. The soil was treated with the test substance and incubated under conditions that allow rapid microbial metabolism. Under these conditions, the available organic carbon in the upper soil is rapidly degraded, resulting in a lack of organic carbon, starvation of microbial cells and resulting in dormancy or budding. If the test is extended beyond 28 days, the sum of these responses is measured in the control (untreated soil) as a loss of microbial metabolic activity. If biomass in carbon-stressed soil is affected by the chemical under the test conditions, it may not recover to the same level as in the control. Therefore, disturbances to the test substance at any time during the test will usually persist until the end of the test.
This test method is mainly used for substances with known environmental concentrations in the soil. For example, the application of pesticides in the soil is known. For agricultural chemicals, only two dose concentrations need to be determined (the concentrations are related to the expected or predicted application rate). Agricultural chemicals can be determined as active ingredients (a, b) or as formulations. However, the test is not only applicable to chemicals with known environmental concentrations, but also to chemicals with unknown concentrations applied to the soil by varying the amount of test substance applied to the soil and the approach to data evaluation. Therefore, for non-agricultural chemicals, the method can be used to determine the effects of different concentrations on carbon transformation. The test results can be used to draw dose-response curves and to calculate the test substance concentration (ECI) value that causes the carbon conversion inhibition percentage in the soil to reach %, where is the carbon conversion inhibition percentage,
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1 Scope
Chemicals
Soil microorganisms
Carbon conversion test
GB/T27855-2011
This standard specifies the test method for the long-term potential effects of a single chemical on the carbon conversion activity of soil microorganisms. This standard is applicable to the evaluation of the toxicity characteristics of the test substances. The carbon conversion test is aimed at the effects of chemicals on the soil microbial flora. 2 Terms, definitions and abbreviations
The following terms, definitions and abbreviations apply to this document. 2.1 Terms and definitions
Carbon conversioncarbon lransformatton
The degradation of organic matter by microorganisms to form the final inorganic carbon dioxide. 2.2 Abbreviations
ECConcentration of the test substance at which the percent inhibition of transformation in soil reaches a% (cffective concentration) ECsConcentration of the test substance at which the percent inhibition of carbon transformation in soil reaches 50% (median ictive concentration) 3 Validity of the test
The evaluation of the test results of agricultural chemicals is based on the difference in the amount of carbon dioxide released or oxygen consumed in the control soil and the treated soil samples. This difference is relatively small (mean ± 25), so large differences in the control group will lead to erroneous results. Therefore, the difference between control parallel samples should be less than ± 15%. 4 Principle
Prepare in advance soil treated with the test substance or untreated soil as a control. If the test substance is an agricultural chemical, it is recommended to use at least two test concentrations, the lowest of which should be related to the highest concentration expected to be used in the field screening. At 0 d, 7 d, 14 d,After 28 days, the treated and control soil samples were mixed with glucose and the glucose-induced respiration rate was measured continuously for 12 hours. The respiration rate was expressed as carbon dioxide released [C (mg)/soil (kg·h) or oxygen consumed [O2 (mg)/soil (kg·h). The average respiration rate of the treated soil samples was compared with that of the control, and the percentage difference in the average respiration rate of the treated soil was calculated based on the control. All tests lasted at least 28 days. If the difference between the treated and untreated soils was equal to or greater than 25% on the 28th day, the test was continued with an interval of 14 days, up to a maximum of 100 days. If the test substance is a non-agricultural chemical, a series of concentrations of the test substance should be added to the soil fill sample and the glucose-induced respiration rate (i.e., the average value of carbon dioxide formation or oxygen consumption) should be measured after 28 days. The results of the series of concentration tests obtained were analyzed by regression method and the EC value, i.e., ECr, EC2s or ECl, was calculated. 5 Test apparatus
The test container should be made of inert material. Appropriate volumes matching the method used for soil culture should be used, i.e., the culture should be carried out as a whole or in a series of individual soil samples. Care should be taken to minimize water loss during the test and to allow gas exchange (for example, the test container can be covered with a polyvinyl film with small holes). When testing volatile substances, sealable or airtight containers should be used. The size of the container should be based on approximately 1/4 of the total volume of the loaded soil sample. To determine glucose-induced respiration, a culture system and instruments for measuring carbon dioxide production or oxygen consumption are required.
6 Test Procedure
6.1 Preparation
6.1.1 Selection of Soil
Use a single soil with the following characteristics: Broken grain content: 50% to 75%,
pH. 5.5 to 7.5;
Organic nitrogen content: 0.5 to 1.5%.
The microbial biomass (12% I) should be determined, and its carbon content should be at least 1% of the total organic carbon in the soil. In most cases, soils with H characteristics represent the worst situation. This is because they have the least absorption of the test chemical and the greatest availability to the microbial flora. Therefore, it is usually not necessary to use other soil tests. However, in some cases, it is expected that the test substance will be used mainly in a specific soil, such as a forest soil with static charges. For chemicals with static charges, another additional soil is used. 6.1. 2. Collection of soil samples
Detailed historical information on the field where the soil to be tested is collected should be available, including: exact location, cover, date of application of crop protection agents, treatment with organic or inorganic fertilizers, biological materials added or accidentally mixed contaminants. The soil selected for sampling should be able to be used for a long time. Pastures with long-term water, fields with cereal crops (except corn) harvested once a year, or densely planted green manure fields are all suitable locations. The sampling site should meet the following requirements: Crop protection agents have not been used for at least one year before sampling. Organic fertilizers have not been applied for at least 6 months. Inorganic fertilizers can be applied only if the crop requires it, and soil samples can be taken at least 3 months after fertilization. Soil treated with pesticides or fertilizers that have biocidal effects (e.g. calcium thiazide) should be avoided. Sampling should be avoided during long-term (more than 30 days) dry or waterlogged periods, or immediately afterwards. The depth of soil samples taken in cultivated land should be 0 ctm to 20 cml. The maximum sampling depth in grassland (pasture) or other types of earth embankments that have not been formed for a long period of time (at least one growing season) can be greater than 20cm (for example 25cm). When transporting soil samples, a storage container should be used, and the temperature conditions should be appropriate to ensure that the original soil properties are not significantly changed. 6.1.3 Storage of soil samples
It is best to use soil freshly collected from the field for testing. If storage is necessary, it should be stored in a dark place where the soil is not frozen for no more than 3 months. During the soil storage period, aerobic conditions should be ensured. If the soil collected comes from an area with at least 3 months of freezing each year, storage at -18℃ for 6 months can be considered. The microbial biomass of the stored soil must be determined before each test. At the same time, the carbon content in the biomass should be greater than 1% of the total organic carbon content of the soil.
6.1.4 Treatment and preparation of test soil fill samples 6.1.4.1 Pre-culture
If stored soil is used, the recommended pre-culture time is The time is 2d to 28d. The temperature and humidity of the soil during the incubation period should be the same as those during the test.
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6.1.4.2 Physical and chemical properties
GB/T27855-2011
Artificially remove coarse debris (e.g., stones, plant residues, etc.) from the soil, and then filter it in a wet state (do not over-dry) to make the particles not too large. Soil moisture should be adjusted with distilled water or deionized water to a water content equivalent to 40% to 60% of the maximum water holding capacity. 6.1.4.3 Preparation of test substances for application to the soil The test substances are generally applied through a carrier. This carrier may be water (for water-soluble substances) or an inert solid substance, such as fine quartz sand (particle size is 0.1mm to 0.5mm). Liquid carriers other than water (organic solvents, such as acetone, tricarbonate, etc.) should be avoided because they can damage the microbial flora. If sand is used as a carrier, the test substance should be dissolved or suspended in the corresponding flux and then applied. In order to obtain the most appropriate distribution of the test substance in the soil, the recommended ratio of sand to soil (dry weight) is: 10 kg sand/kg soil. The control sample is treated with equal amounts of water and quartz sand. When testing volatile chemicals, their volatilization should be avoided as much as possible during the treatment process, and care should be taken to ensure their uniform distribution in the soil (for example, the test substance should be added to the soil from all parts). 6.1.5 Test concentrations
For crop protection agents or other chemicals whose environmental concentrations can be predicted, at least two concentrations should be used. The lower concentration should be set at least to reflect the maximum value that can be predicted to enter the soil under actual conditions, and the higher concentration should be set to an integer multiple of the lower concentration. The concentration of the test substance added to the soil is calculated based on the following: the test substance and the soil are mixed to a thickness of 5 cm and the bulk density of the soil is 1.5. For agricultural chemicals applied directly to soil or chemicals that can be predicted to enter human soil, the recommended test concentration is 5 times the predicted environmental concentration (PEC) and the PEC. For substances expected to be applied to soil multiple times in one season, the test concentration is: the maximum number of expected soil applications multiplied by the predicted environmental concentration. However, the upper limit of the test concentration should not exceed 10 times the single maximum application rate.
If the test substance is not an agricultural chemical, at least 5 concentrations arranged in geometric series should be used. The test concentrations should cover the range required to determine the EC value.
6.2 Test procedures
6.2.1 Treatments and controls
If the test substance is an agricultural chemical, the soil should be divided into 3 equal parts by weight. Two parts are mixed with the carrier containing the test substance, and the other part is mixed with the carrier without the test substance (i.e., the control). Set up at least 3 replicates for each of the treated and untreated soil samples. If the test substance is a non-agricultural chemical, divide the soil into 6 equal parts by weight: 5 of the samples are mixed with the carrier containing the test substance, and the remaining 1 is mixed with a carrier without chemicals. Set up 3 replicates for each of the treated samples or control samples. Careful operation should be taken to ensure that the test substance is mixed and distributed in the treated soil samples. During the mixing process, be careful to avoid compacting or balling the soil. 6.2.2. Incubation of soil samples
Incubation of soil samples can be done in two ways: 1) a whole sample of each treated and untreated soil; 2) a series of samples consisting of individual subsamples of equal size for each treated and untreated soil. When testing for volatiles, only a series of individual subsamples should be used for incubation. When incubating with a whole soil sample, the largest number of treated and untreated soil samples should be prepared, and subsamples can be analyzed as needed during the experiment. The initial number of soil samples prepared for each treatment and control should be determined based on the size of the sample, the number of replicates to be analyzed, and the maximum number of samplings expected. The whole incubated soil should be mixed thoroughly before resampling. When incubating a series of individual soil samples, each treated and untreated whole soil should be divided into the required aliquots and used as needed. When sampling is expected to be performed more than twice in the experiment, aliquots should be prepared to accommodate each sampling and all replicates. At least three replicates of the test soil should be incubated under aerobic conditions. Use suitable containers with sufficient head space in the test to avoid the generation of anaerobic conditions. When testing volatile substances, only a series of separate sub-samples can be used for incubation. 6.2.3 Test conditions and duration
The test should be carried out under dark conditions and at room temperature of ±20℃. During the test, the moisture content of the soil sample should be maintained at 40% to 60% of the maximum water holding capacity of the soil, with a range of ±5%. Distilled water or deionized water can be added when necessary. The test duration is at least 28 days. If the test substance is an agricultural chemical, the carbon dioxide release or oxygen consumption in the treated sample and the control sample should be compared. If the difference between them is greater than 25% on the 28th day, the test should be continued or until the difference between them is equal to or less than 25%, or up to 100 days, whichever is shorter. If the test substance is a non-agricultural chemical, the test should be terminated after 28 days. On the 28th day, the carbon dichloride release or oxygen consumption in the treated and control soil samples should be measured and the EC value calculated. 6.2.4 Soil fill sampling table
If the test substance is an agricultural chemical, the glucose-induced respiration rate in the soil samples should be analyzed at 0d, 7d, 14d and 28d. If a longer test is to be conducted, the measurement should continue every 14 days after 28 days. If the test substance is a non-agricultural chemical, at least 5 test concentrations should be used, and the glucose-induced respiration of the soil samples should be analyzed at the beginning (0d) and the end (28d) of the contact period. If necessary, an intermediate measurement can be added, such as measurement on the 7th day. The EC value of the chemical is determined using the data measured on the 28th day. If necessary, the data of the control sample on the 1st day can also be used to estimate the initial amount of metabolically active microorganisms in the soil.
6.2.5 Determination of glucose-induced respiration rate The glucose-induced respiration rate of each replicate sample of each treatment and control should be determined at each sampling. The soil sample should be mixed with sufficient glucose to rapidly produce a maximum respiration response. The amount of glucose required to produce the maximum respiration response can be obtained by preliminary experiments using a series of glucose concentrations. For sandy soils containing 0.5% to 1.5% organic carbon, 2 000 mg to 4 000 mg of glucose per kg of soil (dry weight) can be used. Grind the glucose with quartz sand until it is powdery and mix it evenly with the soil bag (dry weight) (10 sand/kg soil).
When determining the respiration rate of soil samples adjusted with glucose, whether the measurement is continuous, hourly, or every two hours, the incubator should be kept at a temperature of 20°C ± 2°C. The carbon dioxide release or oxygen consumption should be measured continuously for 12 hours. The measurement should be started as soon as possible, i.e. within 1-2 hours after the addition of glucose. The total carbon dioxide release and the total oxygen consumption should be measured during the 12 hours and the average respiration rate should be determined.
6.3 Quality Control
The difference between parallel control samples should be less than 15 times. 7 Results and Reporting
7.1 Result Processing
7.1.1 Tested Chemicals
If the tested chemical is an agricultural chemical, the carbon dioxide release or oxygen consumption of each parallel soil sample should be recorded and the average value of all replicate samples should be tabulated. The results should be evaluated by appropriate, commonly used statistical methods (e.g., F test, 0.5% significance level). The respiration rate induced by glucose is expressed as: [carbon (ng)/soil (kg·h) or [oxygen (mng)/soil (kg·h)]. The average carbon dioxide generation rate or average oxygen consumption rate of each treated sample is compared with that of the control, and the percentage difference with the control is calculated.
7.1.2 The test substance is a non-agricultural chemical
CB/T 27855-2011
If the test substance is a non-agricultural chemical, the carbon dioxide release and oxygen consumption in each parallel sample are determined, and a drug-response curve is drawn to estimate the EC value. After 28, the respiration rate L caused by the fungicide sugar of the treated sample and the control sample is compared (i.e., carbon (mg)/soil (kg-h) or oxygen (mg)/soil (kg·h)). Using the above data, the inhibition percentage (%) at each test level can be calculated. These percentages are plotted against the logarithm of the concentrations and the EC values and their 95% confidence intervals are calculated using statistical methods [1sI1517]. 7.1.3 Interpretation of results
When evaluating the test results of agricultural chemicals, if the difference in respiration rate between the low concentration treatment (i.e., the maximum expected concentration) and the control is equal to or lower than 25% at any time after 28 days, it can be considered that the product has no long-term effect on carbon transformation in the soil. When evaluating the test results of chemicals other than agricultural chemicals, ECs, EC and (or) ECI values can be used. 7.2 Report The report of results shall include the following information: 1. a) Complete information on the soil tested including: - geographical coordinates of the sampling point (latitude, longitude); - historical information on the sampling point (i.e. vegetation cover, use of crop protectants, use of fertilizers, unexpected pollutants, etc.); - form of use (e.g. agricultural soil, forest, etc.); - sampling depth (cm); - sand/silt/clay content (% dry weight); - pH (in water); - organic carbon content (% dry weight); - nitrogen content (% dry weight); - cation exchange capacity (mmol/kg); - microbial biomass expressed as a percentage of total organic carbon; - reference to the test method used to determine each parameter; complete information on the collection and preservation of soil samples; - soil pre-cultured with bacteria (if applicable). b)
Test substance:
Physical properties + and relevant physico-chemical properties - Chemical identification data including: structural formula, purity (for crop protection agents, the percentage of active ingredients) fluorine content.
c) Test conditions,
Details of soil conditioning with organic substrates; - Concentration values of test chemicals, with appropriate justification for the selected concentration; Details of application of the test substance to the soil: Incubation temperature
- Soil moisture at the beginning and during the test; - Soil incubation method (whole or a series of individual sub-samples) Number of parallel test samples:
- Number of samplings.
The test results should include the following information:
a) the method and equipment used to test the respiratory rate; 5
GB/T27855—2011
Tabular data, including individual values and means of carbon dioxide or oxygen; differences between replicates in treated and control samples, if any corrections were made in the calculation, which should be stated where relevant; the percentage change in respiratory rate caused by respiration at each sampling, or appropriate statement of ECs and their 95% confidence levels, other ECs (i.e. EC25+ or EC10) and their confidence intervals, and plotting of dose-response curves; statistical treatment of the results (stated where appropriate); all information and observations that contribute to the interpretation of the results. References
GB/T 27855—2011
[1] EPPO (19g4). Iecision-Making Scheme for the Environnental Rish Assessment of PlantProtection Chemicals, Chapter 7,Soil Microflara, EPPO Bulletin 1994,24:1-16L2] BBA (1990).Effects on the Activity of thc Soil Microflora. BBA Guidelines for thc OfficialTcsting of Plant Protection Products, VI,1-1 (2nd eds. ),1990[3] EPA (1987). Soil Microbial Community Toxicity Test. EPA 40 CFR Part 797 , 3700. TaxicSubstances Control Act Test Guidelincs; Proposed rulc, Scpternber 28,198?L41 SETAC-Eurupe (1gg5). Procedures for assessing the environmental fate and ecotoxicily ofpenticides,Ed, M, R, Lynch,Pub. SETAC-Europe, BrusselsL5] OECD (1995), Final Report of the (OECD Workshop on Selection of Soils/Sedirments, Belgi-rate,ltaly,1995:18-20
[6l IS0 1038l-6 (1993). Soil guality - Sampling. Guidatce on the collertion, handling and storage ol soil far the assessment of aerobic microbial prucesses in the laboratoryL7] Anderson,JPE (19B7).Handling and Storage of Soils for Pesticide Experiments,in \ Pes-ticide Effects on Soil Microflora\. Eds. L. Sorncrville and MP Greaves,Chap. 3:15-60[8] Anderson,J, PE(1982).Soil Respiration,in Methods of Soil Analysis-Part 2:Chemicaland Microbiological Properties. Agronomy Monograph N° 9. Eds. AL Page, RH Miller and DRKceney.41:831-871
[9] IsO 1l266-1. (1993), Soil Quality - Guidance on Laboratory Tests for Biodegiadatipn inSoil.Partl,AerobicConditions.[10] ISO 14239 (1997E). Soil Quality - Laboratory ineubation systems for measuring the mineralization of organir chemicals in soil undet aerobic conditionsL11] Heinemeye,r O., Insam, H.,Kaiser, E, A, and Walenzik, G. (198g). Soil microbial biomassand respiratiun measurements; an automated technique based on infrared gas analyses. Plant and Soil,116.77-81
[12_ ISO 14240-1 (1997), Soil quality Dctermination of soil microbial biomass Part 1: Sub-strateinduced cspiration method[13] I5O 14240-2 (1997). Soil quality- Determination of soil microbial biomass - Part 2,Fumi-gation cxtraction method
[14] Melkotmes, H. -P, (1986). ( 1949), A sirnplified method al evaluating dose-effert ex-peritnents.Jour.Pharmacol.and Expcr.Thcr.,96:99-113[16] Finney,DJ (1971). Probit Analysis. 3rd ed ,Cambridge,London and New-York[17] Finncy DJ (197$). Statistical Methods in biological A:say ,Griffin,Weycombe,UKFinal Report of the (OECD Workshop on Selection of Soils/Sedirments, Belgi-rate,ltaly,1995:18-20
[6l IS0 1038l-6 (1993). Soil guality - Sampling. Guidatce on the collertion, handling and storage ol soil far the assessment of aerobic microbial prucesses in the laboratoryL7] Anderson,JPE (19B7).Handling and Storage of Soils for Pesticide Experiments,in \Pes-ticide Effects on Soil Microflora\. Eds. L. Sorncrville and MP Greaves ,Chap. 3:15-60[8] Anderson,J, PE(1982).Soil Rcspiration,in Methods of Soil Analysis-Part 2:Chemicaland Microbiological Properties. Agronomy Monograph N° 9. Eds. AL Page, RH Miller and DRKceney.41:831-871
[9] ISO 1l266-1. (1993), Soil Quality - Guidance on Laboratory Tests for Biodegiadatipn inSoil.Partl,AerobicConditions.[10] ISO 14239 (1997E). Soil Quality - Laboratory ineubation systems for measuring the mineralization of organir chemicals in soil undet erobic conditionsL11] Heinemeye,r O. , Insam, H. ,Kaiser,E,A,and Walenzik,G. (198g). Soil microbial biomassand respiratiun measurements; an automated technique based on infrared gas analyses. Plant and Soil,116.77-81
[12_ ISO 14240-1 (1997), Soil quality Dctermination of soil microbial biomass Part 1: Sub-strateinduced cspiration method[13] I5O 14240-2 (1997). Soil quality- Determination of soil microbial biomass - Part 2, Fumi-gation cxtraction method
[14] Melkotmes, H. -P, (1986). Influence of the Amnount of Glucose Added to the Soil on the Effcet of Pesticidcs in Short-Term Respiration, using a Herbicide as an Example ), Nachrichtenbl, Deut. Pllanzenschutzd. ,Braunschweig,38 t 113-120[15] Litchfield.JT and wilcoxon,F. (1949), A sirnplified method al evaluating dose-effert ex-peritnents.Jour.Pharmacol.and Expcr.Thcr.,96:99-113[16] Finney,DJ (1971). Probit Analysis. 3rd ed ,Cambridge,London and New-York[17] Finncy DJ (197$). Statistical Methods in biological A:say, Griffin, Weycombe,UKFinal Report of the (OECD Workshop on Selection of Soils/Sedirments, Belgi-rate,ltaly,1995:18-20
[6l IS0 1038l-6 (1993). Soil guality - Sampling. Guidatce on the collertion, handling and storage ol soil far the assessment of aerobic microbial prucesses in the laboratoryL7] Anderson,JPE (19B7).Handling and Storage of Soils for Pesticide Experiments,in \Pes-ticide Effects on Soil Microflora\. Eds. L. Sorncrville and MP Greaves ,Chap. 3:15-60[8] Anderson,J, PE(1982).Soil Rcspiration,in Methods of Soil Analysis-Part 2:Chemicaland Microbiological Properties. Agronomy Monograph N° 9. Eds. AL Page, RH Miller and DRKceney.41:831-871
[9] ISO 1l266-1. (1993), Soil Quality - Guidance on Laboratory Tests for Biodegiadatipn inSoil.Partl,AerobicConditions.[10] ISO 14239 (1997E). Soil Quality - Laboratory ineubation systems for measuring the mineralization of organir chemicals in soil undet erobic conditionsL11] Heinemeye,r O. , Insam, H. ,Kaiser,E,A,and Walenzik,G. (198g). Soil microbial biomassand respiratiun measurements; an automated technique based on infrared gas analyses. Plant and Soil,116.77-81
[12_ ISO 14240-1 (1997), Soil quality Dctermination of soil microbial biomass Part 1: Sub-strateinduced cspiration method[13] I5O 14240-2 (1997). Soil quality- Determination of soil microbial biomass - Part 2, Fumi-gation cxtraction method
[14] Melkotmes, H. -P, (1986). Influence of the Amnount of Glucose Added to the Soil on the Effcet of Pesticidcs in Short-Term Respiration, using a Herbicide as an Example ), Nachrichtenbl, Deut. Pllanzenschutzd. ,Braunschweig,38 t 113-120[15] Litchfield.JT and wilcoxon,F. (1949), A sirnplified method al evaluating dose-effert ex-peritnents.Jour.Pharmacol.and Expcr.Thcr.,96:99-113[16] Finney,DJ (1971). Probit Analysis. 3rd ed ,Cambridge,London and New-York[17] Finncy DJ (197$). Statistical Methods in biological A:say, Griffin, Weycombe,UKSoil Quality - Laboratory ineubation systems for measuring the mineralization of organir chemicals in soil undet erobic conditions ; an automated technique based on infrared gas analyses. Plant and Soil,116.77-81
[12_ ISO 14240-1 (1997), Soil quality Dctermination of soil microbial biomass Part 1: Sub-strateinduced cspiration method[13] I5O 14240-2 (1997). Soil quality- Determination of soil microbial biomass - Part 2, Fumi-gation cxtraction method
[14] Melkotmes, H. -P, (1986). EinfluB von Glukoscmenge auf die Reaktioh der Kurzzeit-Atmung iltRoden Gegenlber Pflanzenschutzmitteln, Dargestellt am Beispjel eines Herbizide, (Influence of the Amnount of Glucose Added to the Soil on the Effcet of Pesticidcs in Short-Term Respiration, using aHerbicide as an Example), Nachrichtenbl, Deut. Pllanzenschutzd. ,Braunschweig,38 t 113-120[15] Litchfield.JT and wilcoxon,F. (1949), A sirnplified method al evaluating dose-effert ex-peritnents.Jour.Pharmacol.and Expcr.Thcr.,96:99 -113[16] Finney,DJ (1971). Probit Analysis. 3rd ed ,Cambridge,London and New-York[17] Finncy DJ (197$). Statistical Methods in biological A:say, Griffin, Weycombe,UKSoil Quality - Laboratory ineubation systems for measuring the mineralization of organir chemicals in soil undet erobic conditions ; an automated technique based on infrared gas analyses. Plant and Soil,116.77-81
[12_ ISO 14240-1 (1997), Soil quality Dctermination of soil microbial biomass Part 1: Sub-strateinduced cspiration method[13] I5O 14240-2 (1997). Soil quality- Determination of soil microbial biomass - Part 2, Fumi-gation cxtraction methodWww.bzxZ.net
[14] Melkotmes, H. -P, (1986). EinfluB von Glukoscmenge auf die Reaktioh der Kurzzeit-Atmung iltRoden Gegenlber Pflanzenschutzmitteln, Dargestellt am Beispjel eines Herbizide, (Influence of the Amnount of Glucose Added to the Soil on the Effcet of Pesticidcs in Short-Term Respiration, using aHerbicide as an Example), Nachrichtenbl, Deut. Pllanzenschutzd. ,Braunschweig,38 t 113-120[15] Litchfield.JT and wilcoxon,F. (1949), A sirnplified method al evaluating dose-effert ex-peritnents.Jour.Pharmacol.and Expcr.Thcr.,96:99 -113[16] Finney,DJ (1971). Probit Analysis. 3rd ed ,Cambridge,London and New-York[17] Finncy DJ (197$). Statistical Methods in biological A:say, Griffin, Weycombe,UK
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