Metal mining technical guidance: sublethal toxicity checklist, chapter 4


Effluent Sample Identification

Client Name/Location:________________ Collection Approach: Single or Multiple sample(s) (circle one)  

Testing Lab Name/Location:   ___________Test Type: Static-renewal or Flow-through (circle one)

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 “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
Reporting & Method Requirements Reported Data? Met “must” Requirement?
Y N Y N NA
Effluent Sample
Effluent type (eg: process effluent, final effluent etc.)         X
Information on labelling/coding of sample         X
Temperature of sample upon arrival at lab         X
Date for sample collection and date/time for sample receipt at lab         X
Date for test start (within 3 d of sample collection)1          
Test Organisms
Species and common name (Oncorhynchus mykiss - rainbow trout)          
Source (gametes or broodstock obtained from a single population and source)         X
# of female and male broodstock used for fertilization (≥ 4 females; ≥ 3 males)          
Description of procedure for checking sperm motility (milt from ≥ 3 males held in separate vials; milt from each vial examined under microscope 100X; add small amount of water and mix; vials that contain inactive sperm not to be used for fertilization; fresh new milt to be obtained if all vials contain inactive sperm)          
Description (including time interval) of procedure for fertilization of gametes (dry mixing of eggs and milt for a min of 5 min and a max of 20 min)          
Time interval from completion of fertilization until exposure of all groups of eggs to test solutions (test start within 30 min following a period of 5 to 20 min for dry fertilization of eggs)          
Any unusual appearance or treatment of gametes or eggs, before their use in the test         X
Test Conditions and Facilities
Test method (July 1998 2nd edition of EPS 1/RM/28 - E-test)          
Dates or days during test when sub-samples or multiple samples used (ie: days 1-2; days 3-4; days 5-6-7)          
Date for test completion (7 d after fertilization)          
Description of test chamber(s) and associated apparatus (eg: incubation units, pumps etc)          
Person performing test          
Rate of pre-aeration of sample or test solutions, if required, prior to addition of eggs (6.5 ±1 mL/min.L; minimal and controlled)          
Duration of pre-aeration (only if D.O. of test solution < 60% or > 100% upon preparation, in which case pre-aerate sample or all test solutions for 30 min and if necessary for the lesser of an additional period of ≤ 90 min or until 60 - 100% saturation is achieved; test initiated at this point regardless of whether D.O. of 60 - 100% saturation was achieved)          
Rate of aeration (no more than 100 bubbles/min per chamber; minimal and controlled)          
Duration of aeration (if static-renewal test, recommend gentle aeration throughout test; if flow- through test, aerate if necessary to maintain D.O. at 60 to 100% saturation and/or increase rate of exchange; normally aerate during test if effluent)         X
Manner and rate of exchange of test solutions (≥ 0.5 L/g-d; daily renewal of 80% of the old solution in static-renewal test)          
Procedure, if any, for pH adjustment (recommend no adjustment if pH of test solution within 6.5 - 8.5; a second pH-adjusted test might be required/appropriate for pH beyond that range          
Procedure, if any, for sample filtration (recommend no filtration; parallel test with filtered sample is an option)         X
Type(s) and source(s) of control/dilution water (same water used for preparing control and test solutions; adjusted to 14 ± 1°C before use; not supersaturated)          
Sample/subsample adjusted to 14 ± 1°C before use         X
Type and quantity of any chemical(s) added to control/dilution water         X
# and conc. of test solutions (≥ 5 conc. plus a control)          
Volume and depth of solution in each test chamber         X
# of replicates per conc. (equal # of replicates;3 replicates/conc.; 4 recommended)          
# of organisms (equal # of embryos into each test chamber; ≥ 30 / replicate, ie:120 / conc.)          
D.O. and pH of sample just before its preparation and use         X
During the test, Temperature, D.O., and pH (measured in representative concentrations at start and end of each 24-h periods in static-renewal test, or daily in flow-through test; Temperature must be 14 ± 1°C)          
Anything unusual about the test, any deviation from the test method, any problems encountered, any remedial measures taken         X
Results
Average # and % of non-viable embryos (including unfertilized eggs) in each replicate and conc., including control at 7d (test is invalid if mean % viable control embryos is < 70%)          
EC25 (and 95% confidence limits) for embryo viability; indication of quantal statistic employed          
Results of the E test with reference toxicant(s) (conducted concurrently with the effluent test; recommend that the result fall within the warning limits (± 2 SD) of the historic reference toxicant mean)         X
Test with reference toxicant conducted concurrently with E test on effluent?          
Was it conducted under the same experimental conditions?          
Was the test endpoint bracketed by at least 1 test conc. (except for “<0.1% and > 100%”)?          
For the Pulp and Paper Program: Was report submitted within 90 d of test completion?          

* Covers reporting and method requirements outlined in the July 1998 2nd  edition of the early life stages of salmonid fish test and the Amended 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 Sections 7.2 and 7.4 of the Metal Mining EEM Guidance Document and Section 6.3 of the Pulp and Paper EEM Guidance Document.

Page details

Date modified: