Consultant's Report - Minnesota State Legislature

Experimental designs were modified from the veliger toxicity tests conducted by
Fisher et al. (1994) (See Appendix 1). All testing used Costar Inc. (#3516) 6-well plastic
multi-well plates (Corning Inc., Corning, NY), with 10 mL of exposure solution per well.
Tests used six Cu exposure concentrations and a control, with each treatment consisting
of 12 replicates using two well plates. Exposure concentrations* selected for the first and
second experiments were 0.001, 0.002, 0.005, 0.010, 0.020, and 0.050 mg CulL, and for
the third experiment were 0.0001, 0.0005, 0.0010, 0.0050, 0.0100, and 0.0500 mg CulL.
All tests were conducted for 24-h at 18 ± 1°C, the approximate temperature of Lake
Ossawinnamakee in June. Following successful spawning of 2-5 females, the density of
eggs in the combined solution was determined to establish the volume of solution that
contained 150-200 eggs, and this volume was added to each well (i.e., 150-200
eggs/replicate). Eggs were fertilized by the addition of 10 ilL of a concentrated sperm
solution.
• All concentrations listed as nominal.
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