Biological test method for measuring terrestrial plants exposed to contaminants in soil: chapter 7

7.1 Minimum Requirements for a Test-Specific Report
7.2 Additional 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 sorting, preparation, and storage; 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, negative control soil) or coding, as provided to the laboratory personnel;
information on labelling or coding of each sample; and
date of sample collection; date and time sample(s) received at test facility.

7.1.2 Test Organisms

species and source of test seeds;
scientific name, seed variety, and lot number; and
any unusual appearance or treatment of the seeds, 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 seeds per test vessel and treatment;
dates when test was started and ended;
measurements of light intensity adjacent to surface of soil in test vessels;
for each soil sample--any measurements of soil particle size, moisture content, water-holding capacity, pH, and conductivity; and
for each composite sample of subsamples taken at the same time from all replicates of each treatment--all measurements of temperature, pH, moisture content, and water-holding capacity.

7.1.6 Test Results

number of seedlings and observations on seedling condition in each test vessel, as noted during each observation period over the test duration;
mean (± SD) percent emergence in each treatment, including control(s), on Day 7 (if determined) and at test end (Day 14 or Day 21, depending on species of test organism);
mean (± SD) shoot length of individual plants surviving in each treatment [(including the control(s)] at test end; mean (± SD) root length of individual plants surviving in each treatment at test end; mean (± SD) shoot dry weight of individual plants surviving in each treatment at test end; mean (± SD) root dry weight of individual plants surviving in each treatment at test end;
mean (± SD) shoot and root wet weight of individual plants surviving in each treatment [(including the control(s)] at test end, if determined;
any EC50 (including the associated 95% confidence limits, and if calculated, the slope) determined; any additional ECx (e.g., EC20) calculated;
any ICp (together with its 95% confidence limits) determined for the data on growth (i.e., shoot and root lengths and shoot and root wet and dry weights of individual plants surviving 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- or 10-day (depending on test species) ICp (including its 95% confidence limits) performed with the reference toxicant in conjunction with the definitive soil toxicity test, using the same lot of test seed; geometric mean value (± 2 SD) for the same reference toxicant and test species, as derived at the test facility in previous 7- or 10-day ICp 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 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 and address of seed supplier;
year of collection (if applicable), packet size, lot number, percent germination rating, date of germination rating, date of purchase, name of supplier, and date seed package was opened;
description of procedures used to sort seeds; and
description of storage conditions and procedures, including temperature and duration of seed-lot storage.

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 (if artificial soil) or pretreatment (if natural soil) of negative control soil;
source of natural soil; history of past use and records of analyses 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

procedure for adding seed to test vessels;
relative humidity of test facility;
appearance of each sample (or mixture thereof) in test vessels; changes in appearance noted during test;
records of hydration of test soils in each test vessel throughout duration of test and qualitative description of drying of soils during the test (e.g., rate, appearance);
any other physicochemical measurements (e.g., analyses of aliquots from the same batch to determine conductivity, homogeneity, contaminant concentration, total volatile solids, biochemical oxygen demand, chemical oxygen demand, total inorganic carbon, cation exchange capacity, oxidation-reduction 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 pore water;
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;
percent emergence of plants in each test vessel at test end (Day 14 or Day 21); and on Day 7 (if determined);
number of surviving plants in each test vessel at test end (Day 14 or Day 21); mean individual shoot/root length; and replicate dry weight and mean individual dry weights of shoots and roots (and the same for shoot and root wet weight, if performed) of plants surviving in each test vessel at test end; for regression analyses, hold on file 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 charts (for ICps causing reduced shoot lengths) showing the most recent and historic results for toxicity tests with the reference toxicant and the selected species of test organism used in these tests;
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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Date modified:: 2017-06-13

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