Biological test method for measuring survival of springtails exposed to contaminants in soil: chapter 7


Section 7

Reporting Requirements

Each test-specific report must indicate if there has been any deviation from any of the must requirements delineated in Sections 2 to 6 and, if so, provide details of the deviation(s). The reader must be able to establish from the test-specific report whether the conditions and procedures preceding and during the test rendered the results valid and acceptable for the use intended.

Section 7.1 provides a list of items that must be included in each test-specific report. A list of items that must either be included in the test-specific report, provided separately in a general report, or held on file for a minimum of five years, is found in Section 7.2. Specific monitoring programs, related test protocols or regulations might require selected test-specific items listed in Section 7.2 (e.g., details about the test material and/or explicit procedures and conditions during sample collection, handling, transport and storage) to be included in the test- specific report, or might relegate certain test-specific information as data to be held on file.

Procedures and conditions common to a series of ongoing tests (e.g., routine toxicity tests for monitoring or compliance purposes) and consistent with specifications in this document, may be referred to by citation or by attachment of a general report that outlines standard laboratory practice.

Details on the procedures, conditions and findings of the test, which are not conveyed by the test-specific report or general report, must be kept on file by the laboratory for a minimum of five years so that the appropriate information can be provided if an audit of the test is required. Filed information might include:

  • a record of the chain-of-continuity for field-collected or other samples tested for regulatory or monitoring purposes;
  • a copy of the record of acquisition for the sample(s);
  • chemical analytical data on the sample(s) not included in the test-specific report;
  • bench sheets for the observations and measurements recorded during the test;
  • bench sheets and warning chart(s) for the reference toxicity tests;
  • detailed records of the source of the test organisms, their taxonomic confirmation and all pertinent information regarding their culturing and/or holding and acclimation and health; and
  • information on the calibration of equipment and instruments.

Original data sheets must be signed or initialled, and dated by the laboratory personnel conducting the tests.

7.1 Minimum Requirements for a Test-specific Report

The following items must be included in each test-specific report.

7.1.1 Test Substance or Material

  • brief description of sample type (e.g., waste sludge, reference or contaminated field-collected soil, horizon, negative control soil) or coding, as provided to the laboratory personnel;
  • information on labelling or coding of each sample;
  • information on sample horizons as they were collected (i.e., number, relative depth of each soil horizon), for test, reference and negative control soils, if applicable; and
  • date of sample collection; date and time sample(s) received at test facility.

7.1.2 Test Organisms

  • species and source of brood stock and test organisms;
  • age-range of F. candida, O. folsomi, F. fimetaria or P. minuta at start of test; and
  • any unusual appearance, behaviour or treatment of the organisms, before their use in the test.

7.1.3 Test Facilities

  • name and address of test laboratory; and
  • name of person(s) performing the test (or each component of the test).

7.1.4 Test Method

  • citation of biological test method used (i.e., as per this document);
  • design and description if specialized procedure(s) (e.g., preparation of mixtures of spiked soil; preparation and use of solvent and, if so, solvent control) or modification(s) of the standard test method described herein;
  • brief description of frequency and type of all measurements and all observations made during test; and
  • name and citation of program(s) and methods used for calculating statistical endpoints.

7.1.5 Test Conditions and Procedures

  • design and description of any deviation(s) from, or exclusion of, any of the procedures and conditions specified in this document;
  • number of discrete samples per treatment; number of replicate test vessels for each treatment; number and description of treatments in each test including the control(s); test concentrations (if applicable);
  • volume and/or mass of soil in each test vessel;
  • number of organisms per test vessel and treatment;
  • dates when test was started and ended;
  • feeding regime and ration during the test;
  • for each soil sample, any measurements of soil particle size, moisture content, water-holding capacity, pH and conductivity; and
  • for each composite sample of subsample taken at the same time from all replicates of each treatment, all measurements of temperature (air and soil), pH, moisture content and water-holding capacity.

7.1.6 Test Results

  • mean (± SD) percent survival of adult Collembola in each treatment, including control(s) and reference soils on Day 21 for F. fimetaria and P. minuta, and Day 28 for F. candida and O. folsomi; mean (± SD) number of surviving juveniles in each treatment, including control(s) and reference soils on Day 21 for F. fimetaria and P. minuta, and Day 28 for F. candida and O. folsomi;
  • any LC50 (including the associated 95% confidence limits and, if calculated, the slope) determined; any additional LCp (e.g., LC25) calculated;
  • any ICp (together with its 95% confidence limits) determined for the data on reproductive success (i.e., number of surviving juvenile springtails in each treatment at test end); details regarding any transformation of data, and indication of quantitative statistical method used or procedures applied to the data;
  • for a multi-concentration test with chemical-spiked soil, indication as to whether results are based on nominal or measured concentrations of chemical(s) or chemical product(s); all values for measured concentrations;
  • results for any 7-day LC50 for O. folsomi, or F. fimetaria, or 14-day LC50 for F. candida and P. minuta (including its 95% confidence limits) performed with the reference toxicant in conjunction with the definitive soil toxicity test; geometric mean value (± 2 SD) for the same reference toxicant and test species, as derived at the test facility in previous 7- or 14-day LC50 tests using the procedures and conditions for reference toxicity tests described herein; and
  • anything unusual about the test, any problems encountered, any remedial measures taken.

7.2 Additional Reporting Requirements

The following list of items must be either included in the test-specific report or the general report, or held on file for a minimum of five years.

7.2.1 Test Substance or Material

  • identification of person(s) who collected and/or provided the sample;
  • records of sample chain-of-continuity and log-entry sheets; and
  • conditions (e.g., temperature, in darkness, in sealed container) of sample upon receipt and during storage.

7.2.2 Test Organisms

  • name of person(s) who identified the organisms and the taxonomic guidelines used to confirm species;
  • history and age of brood stock, for any culture used to provide test organisms;
  • description of culture conditions and procedures, including temperature, lighting, type and amount of substrate and details on its periodic renewal, measurements of substrate quality, density of springtails, feeding regime and quantity, records of health and performance indices;
  • procedures used to count, handle, sort and transfer animals; and those to determine their mortality, condition, appearance and behaviour; and
  • source and composition of food, procedures used to prepare and store food, feeding method(s), feeding frequency and ration.

7.2.3 Test Facilities and Apparatus

  • all results for initial tests with negative control soil and reference toxicant, undertaken by the laboratory previously inexperienced with performing the biological test method described herein in advance of any reporting of definitive test results (see Section 3.2.1);
  • description of systems for providing lighting and for regulating temperature within test facility;
  • description of test vessels and covers; and
  • description of procedures used to clean or rinse test apparatus.

7.2.4 Negative Control Soil or Reference Soil

  • procedures for the preparation (ifartificial soil) or pretreatment (if natural soil) of negative control soil;
  • source of natural soil; history of past use and records of analysis for pesticides or other contaminants;
  • formulation of artificial soil, including sources for the constituents and conditions and procedures for hydration and pH adjustment; and
  • storage conditions and duration before use.

7.2.5 Test Method

  • procedures used for mixing or otherwise manipulating test soils before use; time interval between preparation and testing;
  • procedure used in preparing stock and/or test solutions of chemicals; description and concentration(s) of any solvent used;
  • details concerning aliquot sampling, preparation, and storage before physicochemical analysis, together with available information regarding the analytical methods used (with citations); and
  • use and description of preliminary or range-finding test.

7.2.6 Test Conditions and Procedures

  • measurements of light intensity adjacent to surface of soil in test vessels;
  • procedure for adding test organisms to test vessels;
  • appearance of each sample (or mixture thereof) in test vessels; changes in appearance noted during test;
  • records of the addition of test water on the surface of the soil in each test vessel throughout the test, for increasing moisture content;
  • any other physicochemical measurements (e.g., analyses of aliquots from the same batch to determine homogeneity; contaminant concentration, total volatile solids, biochemical oxygen demand, chemical oxygen demand, total inorganic carbon, cation exchange capacity, redox potential, total nitrogen) made before and during the test on test material (including negative control soil and reference soil) and contents of test vessels; including analyses of whole soil and porewater;
  • any other observations or analyses made on the test material (including samples of negative control soil or reference soil); e.g., qualitative and/or quantitative data regarding indigenous macrofauna or detritus, or results of geochemical analyses; and
  • any chemical analyses of the concentration of chemical in stock solution(s) of reference toxicant and, if measured, in test concentrations.

7.2.7 Test Results

  • results for any range-finding test(s) conducted;
  • number of surviving adult springtails in each test vessel at test end (i.e., Day 21 for F. fimetaria and P. minuta, and Day 28 for F. candida and O. folsomi); number of surviving progeny in each test vessel at test end (i.e., Day 21 or Day 28); for regression analyses, information indicating sample size (e.g., number of replicates per treatment), parameter estimates with variance, any ANOVA table(s) generated, plots of fitted and observed values of any models used, and the output provided by the statistical program (e.g., SYSTAT);
  • warning chart showing the most recent and historic results for acute toxicity tests with the reference toxicant and the species of test organisms used in these tests;
  • results for any 21- or 28-day chronic test(s) performed with a reference toxicant;
  • graphical presentation of data; and
  • original bench sheets and other data sheets, signed and dated by the laboratory personnel performing the test and related analyses.
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