Metal mining technical guidance: sublethal toxicity checklist, chapter 1
Effluent Sample Identification
Client Name/Location:_________________________
Testing Lab Name/Location:_________________________
Instructions for Completion of Checklist_________________________
- Column one of the table lists reporting & method requirements (as per 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) | |||||
| Test Organisms | |||||
| Species (Pimephales promelas) | |||||
| Source (all from the same batch) | |||||
| Age at start of test (≤ 24 h old larvae) | |||||
| Any unusual appearance, behaviour, or treatment of larvae before their use in the test (fish appearing abnormal not selected for test) | X | ||||
| Brief statement confirming inflated swimbladders (larvae must not be used if their swimbladders are not inflated) | |||||
| Health Criteria for Test Organisms Cultured at Laboratory | |||||
| Weekly % mortality for breeding stock, up to and including 7-d period preceding collection of eggs for test, monitored and recorded for at least 5 days of the week (mortality < 5% of the general population being reared; mortality < 5% of the fish in individual aquaria; if mortality between 5 and 10%, holding of fish extended for at least another 7d before collection of eggs, until less than than 5% mortality in 7d is realized; if combined incidence of mortalities and diseased fish in adult breeding stock > 10% per week at any time, that stock of fish must not be used to produce test fish) | |||||
| Test Organisms Imported from External Supplier (EC, 1999) | |||||
| Source (name and address of supplier, all from the same source) | |||||
| Each shipment includes statement identifying age of test organisms, date and time of shipment, # of organisms shipped | |||||
| Any unusual appearance, behaviour, or treatment of larvae by the supplier prior to shipping, or by the testing laboratory upon arrival at the testing laboratory or in the period immediately preceding the test (excessive crowding of larvae during shipment must be avoided (i.e., such that stress is minimized and oxygen deficiency prevented); test organisms must be disease free, appear healthy (not discoloured or otherwise stressed), and behave normally; test organisms recovered from disease or previous test exposure must not be used in a test) | |||||
| Temperature and D.O. of water in which organisms are shipped just before being shipped and upon arrival at the testing laboratory (temperature change ≤ 3°C per 24 hours during transport; D.O. maintained at ≥ 80% during transport) | |||||
| Brief description of test organism acclimation rate and procedure at the testing laboratory (test organisms must not be stressed during acclimation) | |||||
| Any deviations from test-method-specific “must” requirements for culturing of test organisms, facilities and apparatus used for culturing test organisms, and culture/holding-water conditions | X | ||||
| Health Criteria for Test Organisms Imported from External Supplier (EC, 1999) | |||||
| Weekly % mortality for breeding stock, up to and including 7-d period preceding collection of eggs for test, recorded for at least 5 days of the week (mortality < 5% of the general population being reared; mortality < 5% of the fish in individual aquaria; if mortality between 5 and 10%, holding of fish extended for at least another 7d before collection of eggs, until no more than 5% mortality in 7d is realized; if mortality of adult breeding stock > 10% per week at any time, that stock of fish not be used to produce test fish) | |||||
| Test organism mortality rates (%) upon arrival at the test laboratory and in the 24-hour period preceding a test (mortality ≤ 10% in 24-hour period preceding the test) | |||||
| Test Conditions and Facilities | |||||
| Test method cited (EPS 1/RM/22, second edition February 2011) | |||||
| Dates or days during test when subsamples or multiple samples used (i.e., days 1-2; days 3-4; days 5-6-7) | X | ||||
| Date for test completion (test ended on day 7) | |||||
| Description of test vessels (size, shape, type of material) | X | ||||
| Person performing test | |||||
| 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 test with pH adjusted solution) | X | ||||
| Procedure, if any, for sample filtration or settling and decanting (recommend none) If indigenous organisms present, filter through a sieve with 60µm mesh openings before use) | |||||
| Procedure, if any, for hardness adjustment (recommend none) | X | ||||
| 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 ≥ 250 mL) | |||||
| # of replicates per conc. (equal # of replicates; ≥ 3 replicated / conc. and control) | |||||
| Second control using lab water supply to be set-up if water other than lab water supply normally used to culture the breeding stock is used as control/dilution water (was second control set up?) | |||||
| # of organisms (≥ 10 fish/test vessel; equal # of fish among treatments) | |||||
| D.O. and pH of sample just before its use | X | ||||
| Temperature, D.O. and pH of test solutions and controls; test solution (> 80%) renewed at 24 hr intervals for test duration (measured at least at start (fresh solutions) and end (old solutions) of each 24 h exposure in representative conc. (control, low, med, high); 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 | |||||
| Swimming behaviour, # and % mortality in each replicate test vessel (including control) as noted during each 24-h observation period over the 7 days of exposure (test is invalid if control mortality > 20%) | |||||
| Mean (±SD) % mortality for each treatment at test end | |||||
| Combined and cumulative mean (±SD) % of control fish which either appear dead, moribund, displayed loss of equilibrium, or showed clearly atypical swimming behaviour, at each period of observation including at test end (test is invalid if > 20%) | |||||
| Average dry weight per surviving larvae from control treatment at test end (test is invalid if < 250 µg) | |||||
| Mean (±SD) biomass for each treatment including control(s) at the end of the test, as used for the ICp calculation (i.e., total weight of surviving fish divided by number of fish at test start, presumably 10) | |||||
| If receiving water used as control/dilution, then growth, mortality and swimming behaviour of fish in the lab control water was compared to that in the sample of receiving water | X | ||||
| LC50 (and 95% confidence limits) for survival; indication of quantal statistic employed | |||||
| IC25 (and 95% confidence limits) for biomass; 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 transformation of the data, more than one model tried to fit data and best fit chosen | |||||
| Endpoints generated by regression analysis are bracketed by test concentrations (i.e., extrapolation is not acceptable, except for “> 100 %”) | |||||
| Data from any high test concentration(s) resulting in 0 surviving larvae for all replicates must be removed prior to regression analyses | |||||
| 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) Logarithm of concentration must be used in all calculations of mean and standard deviation. | |||||
| If in-house cultured organisms are used, reference toxicant test conducted at the same time or within 14 d of the start of the effluent test (if performed at same time the same batch of test organisms used) | |||||
| If test organisms are imported, the reference toxicant test must be conducted on the same batch of larvae at the same time as the effluent test | |||||
| The ref tox test must be conducted under the same experimental conditions (description of any deviations from test method) | |||||
| EEM Program Specific Requirements | |||||
| If applicable, statement that EC guidance document on the importation of test organisms has been followed (EC, 1999) | |||||
| 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%”)? | - | - | |||
* Covers reporting and method requirements outlined in the test of growth and survival using larval fathead minnows, second edition published in 2011, and Schedule 5 of the Metal Mining Effluent Regulations (June 2002)
EC (1999), Environment Canada “Recommended Procedure for the Importation of Test Organisms for Sublethal Toxicity Testing”, unpublished report, 22 p.
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