Metal mining technical guidance: sublethal toxicity checklist, chapter 6
Effluent Sample Identification
Client Name/Location: __________________
Testing LabName/Location: __________________
Instructions for Completion of Checklist
- Column one of the table lists reporting & method requirements (including the Regulations Amending the Pulp and Paper Effluent Regulations and Schedule 5 of the Metal Mining Effluent Regulations). Reporting requirements are specified in regular type, while method “must” requirements are indicated in bold type.
- In column two of the table, mark under the Y if data have been reported, or under the N if data have not been provided as required by the minimum reporting of the test result.
- For column 3, this reported information can be either method minimum reporting or an EEM program requirement which is needed to evaluate whether “must” requirements have been met. If data meet the “must” requirements specified, mark under the Y in the third table column or under the N if it is not the case. Items which have no associated method or EEM “must” requirement have been hard-coded with an X under the NA.
| Reporting & Method Requirements and EEM Requirements | Reported Data? | Met “must” Requirement? | |||
|---|---|---|---|---|---|
| Y | N | Y | N | NA | |
| Effluent Sample | |||||
| Effluent type identified (e.g., process effluent, final effluent) | X | ||||
| Information on labelling/coding of sample | X | ||||
| Measurement of temperature of the sample or of one subsample upon receipt at the laboratory | X | ||||
| Date(s) for sample collection and date(s)/time(s) for sample receipt at lab | X | ||||
| Date for test start (≤ 3 d of sample collection)1 | |||||
| Test Organisms | |||||
| Species (Ceriodaphnia dubia) | |||||
| Source (cultures from a single brood organism to provide test organisms) | |||||
| Age at start of test (< 24 h old neonates; all within 12 h of the same age) | |||||
| Any unusual appearance, behaviour, or treatment of test organisms, before their use in the test |
X | ||||
| Health Criteria | |||||
| Mean % mortality of the brood organisms in individual cultures during 7-d period preceding test (no more than 20% mortality) | |||||
| Mean # of surviving young produced per adult in individual cultures during 7-d period preceding test (average of at least 15 young per adult during their first 3 broods) | |||||
| Neonates used to start a test must be taken from individual brood cultures containing at least 8 young produced during the 3rd or subsequent brood | |||||
| Ephippia must not be present in the culture | |||||
| Test Conditions and Facilities | |||||
| Test method (EPS 1/RM/21, 2nd edition, February 2007) | |||||
| Test solution renewed at ≤24 h intervals for test duration | |||||
| Dates or days during test when sub-samples or multiple samples used (i.e., days 1-2; days 3-4; days 5-6-7) | |||||
| Date for test completion (when 60% of control organisms have 3 broods or within 8 days, whichever occurs first) | |||||
| Description of test vessels (size, shape, type of material) | X | ||||
| Person performing test | X | ||||
| Rate of pre-aeration of solutions, if required, prior to addition of test organisms (not exceeding 100 bubbles/min per test vessel; minimal and controlled) | |||||
| Duration of pre-aeration (only if D.O. of test solution < 40% or > 100% upon preparation, in which case pre-aerate all solutions for the lesser of 20 min and attaining 40% of air saturation in the highest test conc.; test initiated at this point regardless of whether D.O. of 40 - 100% saturation was achieved) | |||||
| Rate/duration of aeration, if any, during exposure of test organisms (normally no aeration; not exceeding 100 bubbles/min per test vessel; minimal and controlled) | |||||
| Procedure, if any, for pH adjustment (recommend no adjustment if pH of test solution within 6.5 - 8.5; pH adjusted outside this range is an option or parallel testing with and without pH adjusted solution) | X | ||||
| Procedure, if any, for sample filtration (recommend none; if indigenous organisms, filter through a sieve with 60 µm mesh openings before use) or hardness adjustment (recommend none) | |||||
| Type(s) and source(s) of control/dilution water (same water used for preparing control and test solutions; adjusted to 25 ± 1°C before use; not supersaturated) | |||||
| Type and quantity of any chemical(s) added to control/dilution water | X | ||||
| Sample / subsample adjusted to 25 ± 1°C before use | |||||
| # and conc. of test solutions (≥ 7 test conc. plus a control) | |||||
| Volume and depth of solution in each test vessel (volume ≥ 15 mL) | |||||
| # of replicates per conc. (equal # of replicates; ≥ 10 replicates/conc.) | |||||
| Second control using culture water source set-up if water other than that in which organisms have been cultured is used as control/dilution water (was second control set up?) | |||||
| # of organisms (1 neonate/test vessel; equal # of neonates among treatments) | |||||
| D.O. and pH of sample just before its use | X | ||||
| Temperature, D.O. and pH of test solutions and controls (measured at least at start and end (before renewal of solutions) of each 24 h exposure in representative conc.; temperature 25 ± 1°C | |||||
| Anything unusual about the test, any deviation from the test method, any problems encountered, any remedial measures taken | X | ||||
| Test Endpoints and Calculations | |||||
| Mean % mortality for first-generation daphnids in each test solution (including control) at each period of observation and test end (test is invalid if control mortality > 20%at any time) | |||||
| Mean # of neonates (+ SD) per first-generation daphnid in each test solution (including control), during its first three broods only (test is invalid if 3 broods not produced in ≥ 60% of controls within 8 d and if an average of < 15 live young produced per surviving adult in the controls upon ≥ 60% of the adults achieving their 3rd brood. Note: 2 or more neonates constitutes a brood, if 1 neonate is produced it is scored as part of the neonate count for the previous or following day, any neonates produced as a 4th or subsequent brood are not included in the total # of neonates for any treatment) | |||||
| If receiving water used as control/dilution water, mortality and reproduction rates for C. dubia held in 10 replicate solutions of culture water compared to those for test organisms held in the 10 replicate solutions of receiving water | |||||
| LC50 (and 95% confidence limits) for survival; indication of quantal statistic employed | |||||
| IC25 (and 95% confidence limits) for reproduction; indication that regression analysis was used (ICPIN can still be used to derive an ICp if the data do not allow regression statistics); program name & citation of statistical method(s) used; details regarding any weighting techniques applied to the data | |||||
| Endpoints generated by regression analysis are bracketed by test concentrations (i.e., extrapolation is not acceptable) | |||||
| Outliers (if any) are identified and their removal justified | X | ||||
| Results and duration of the reference toxicant test(s) (recommend that the result fall within the warning limits (± 2 SD) of the historic reference toxicant mean) | |||||
| Was a reference toxicant test conducted within 14 d before or after the Ceriodaphnia test on effluent was started or during it? | |||||
| Was the ref tox test conducted under the same experimental conditions? | |||||
| EEM Program Specific Requirements | |||||
| Were quantitative endpoints for effluent and reference toxicant tests provided (i.e., no less than values reported)? | - | - | |||
| Were the test endpoints bracketed by at least one test concentration (except for “>100%”)? | - | - | |||
| For the Pulp & Paper EEM Program: was the report submitted within 90 d of test completion? | |||||
* Covers reporting and method requirements outlined in the test of reproduction and survival using the cladoceran Ceriodaphnia dubia, amended in 2007, and Regulations Amending the Pulp and Paper Effluent Regulations, and Schedule 5 of the Metal Mining Effluent Regulations (June 2002)
1 For more information on test start date, please refer to Section 7.4 of the Metal Mining EEM Guidance Document and Section 6.3 of the Pulp & Paper EEM Guidance Document.
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