BS ISO 21285:2019 pdf free download.Soil quality Inhibition of reproduction of the soil mite (Hypoaspis aculeifer) by soil contaminants.
1 Scope
BS ISO 21285 specifies a chronic test method for evaluating the habitat function of soils and determining effects of soil contaminants and substances on the reproduction of Hypauspis oculeifer by
— mainly — alimentary uptake. This method is applicable to soils and soil materials of unknown quality, e.g. from contaminated sites, amended sells, soils alter remediation, industrial, agricultural or other sites under concern and waste materials (e.g. dredged material, municipal sludge from a wastewater treatment plant, composed material, or manure, especially those for possible land disposal). The reproduction (= number of juveniles) Is the measured parameter of the test. The test reflects the bloavailability of a mixture of contaminants In natural soils (contaminated site soils) to a species which represents a trophic level which is not covered by other ISO standards. This test is not intended to repLace the earthworm (see 150 1 1268.2) or Collembola (see ISDJ.1262) reproduction tests since this species belongs not only to a different trophic group but also a different taxonomic group (. mites; I.e. arachnids) than those used usually.
Effects of substances are assessed using a standard soil, preferably a defined artificial soil substrate, For contaminated soils, the effects are determined in the soil to be tested and in a control soil. Depending on the objective of the study, the control and dilution substrate (dilution series of contamhiated soil) are either an uncontaminated soil comparable to the soil to he tested (reference soil) or a standard soil (e.g. artificial soil).
This document provides information on how to use this method for testing samples (soils or substances) under temperate conditions.
BS ISO 21285 is not applicable 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.
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.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content constitutes requirements of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.
ISO 10390. SoIl quality — Determination of p11
ISO 10694. Soil quality — Determination of organic and total carbon after dry combustion (elementary analysis)
ISO 11260. Soil quality — Determination of effective cation e.wchange capacity and base saturation level using barium chloride solution
ISO 11277. Soil quality — Determination of particle srze distribution in mineral soil material — Method by sieving and sedimentation
ISO 11465. Soil quality — Determination of dry matter and water content on a moss basis — Gravimetric method
ISO 18400-206. SoIl quality — Sampling — Part 206: CollectIon, handling and storage of soil under aerobic conditions for the assessment of microbiological processes, biomass and diversity in the laboratory
— Sphagnum peat finely ground Ia particle size of (2 ± 1) mm Is 5% acceptable) and with no visible plant remains
— Kaolinite clay containing not less than 30% kaolinlte 20%
— Industrial quartz sand (dominant fine sand with more than 50% 74% of particle size 0,05 mm to 0.2 mm)
Approximately 0,3 % to 1.0% calcium carbonate (CaCO3, pulverised, analytical grade) are necessary to get a pH of 6,0± 0,5.
NOTE) Taking the properties of highly non-polar (log K > 2) or ionizing substances into account. S % of peat have proven to be sufficient for maintaining the desired structure olthe artifIcial soil.
NOTE 2 It has been demonstrated that Hypoaspis acuklfer can comply with the validity criteria even on reproduction when tested in field soils with lower organic carbon content (e.g. 2,7 %). and there is experience that this can be achieved in artificial soil with 5% peat. Therefore. Ills not necessary before using such a soil in a definitwe test to demonstrate the suItability olihe arflfictal soil for allowing the test to comply with the validity criteria unless the peat contents lowered more than specIfied above.
Prepare the artificial soil at least three days prior to start the test, by mixing the dry constituents listed above thoroughly in a Iarge•scale laboratory mixer. A portion of the deionized water required is added while mixing is continued. Allowance should be made for any water that is used for introducing the test substance into the soil. The amount of calcium carbonate required can vary, depending on properties of the individual batch of sphagnum peat and should be determined by measuring sub-samples immediately before the test (see Annex C). Store the mixed artificial soil at room temperature for at least two days to equilibrate acidity. To determine pH and the maximum water holding capacity, the dry artificial sell is pre-moistened one or two days before starting the test by adding delonised water to obtain approximately half of the required final water content of 40% to 60% of the maximum water holding capacity.
The total water-holding capacity shall be determined in accordance with Annex.B. the pH shall be determined according to 150J0320.
5.3 Reference substance
5.3.1 General. To ensure the quality of the test system, tests should be performed regularly (once or twice a year) with a reference substance.
The NOEC and/or the EC1 of a reference substance shall be determined to provide assurance that the laboratory test conditions are adequate and to verify that the response of the test organisms did not change over time. The reference substance can be tested in parallel to the determination of the toxIcity of each test sample at one concentration, which needs be demonstrated beforehand in a dose response study to result in an effect of about 50 %. In this case, the number of replicates should be the same as that in the controls. Alternatively, the reference substance is tested once or twice a year in a dose- response test. Depending on the design chosen, the number of concentrations and replicates and the spacing factor differ (see Z.L3). but a response of 10% to 90% effect should be achieved (spacing factor of 1,8). Dimethoate as well as boric acid are suitable reference substances that have shown to affect reproductionl25l.
The EC50 for dimethoate based on the number of juveniles should fall in the range between 3,0 mg a.s. (active substance)/kg soil (dry mass) and 7.0 mg a.s. (active substance)/kg soil (dry mass). Based on the results obtained with boric acid so far, the ECo based on the number of juveniles should tail in the range between 100 mg/kg (dry mass) soil and 300 mg/kg (dry mass) soil.
Therefore, in cases where various dilutions (concentrations) of each sample of field-collected soil with negative control soil are tested, data are preferably analysed by the ER/EC1-approach or the NOEC-approach.
— ER5/EC5 (effect concentration)-approach.
The ER/EC approach can only be used 11 a clear dose response relationship is found. Wherever possible, the R2 should be 0.7 or higher and the test mixtures used encompass 20 % to 80 % effects. If these requirements are not fulfilled, expert knowledge is necessary for the interpretation of the test results.
To compute an ERx/ECx-value. the treatment means are used for regression analysis after an appropriate dose-response function has been found (e.g. probit or logistic function). A desired ERx/ECx is obtained by inserting a value corresponding tox% of the control mean into the formula found by regression analysis. Since ECg0 values have smaller conlidence limits compared with smaller effect concentrations (e.g. ER2O/EC2O), it is recommended to determine ERo/ECo values.
— NOEC (No-observed-effect-concentration)-approach.
First, a statistical analysis of the homogeneity olthe variances shall be made. e.g. by using Cochran’s test. With homogeneous data, an appropriate statistical analysis, e.g. a One-Way Analysis of Variance (ANOVA). followed by a one-sided Punnett test (a O,OS), should be performed. If the homogeneity requirement is not fulfilled, it is recommended to evaluate If an appropriate transformation of the data can solve the problem. Otherwise non-parametric methods, e.g. the U-test by Mann & Whitney or the Bonferronl-U’test can be used.
If a limit test has been performed and the pre-requisites (normality, homogeneity) of parametric test procedures are fulfilled, the Student-t-test. otherwise the unequal-variance t-test (Welch t-test) or a nonparametric test, such as the Mann-Whitney-U-test may be used.
In any case the results of the statistical evaluation shall be biologically Interpreted.
11 Test report
The test report shall include the following Information:
a) a reference to this document, I.e. ISO 21285:
b) the results, expressed as In 6.Z
c) detailed description of the test sample and Information on physical and chemical properties if helpful for the interpretation of the test result; similarly, detailed information on the soils used Is needed:
— the origin of the field soil used as a control and dilution soil (if appropriate);
— n case of testing soil material: a table with results of chemical analyses of the tested soil (e.g. including C,. heavy metal contents), if available;
d) complete description of the biological material employed (species, age, breeding conditions, supplier);
e) method of preparation of the test sample together with an indication of the auxiliary substances used for a low-/non-water-soluble substance:
f) results obtained with the reference substance;
g) detailed conditions aithe test environment;
h) a table giving the percent mortality of adults at each concentration and in the control(s);
I) number of dead or missing adults and number o(olTsprlng per test container at the end of the test.