Supplemental Data for
Chronic Sensitivity of White Sturgeon (Acipenser transmontanus) and Rainbow
Trout (Oncorhynchus mykiss) to Cadmium, Copper, Lead, or Zinc in Laboratory
Water-Only Exposures
Ning Wang *†, Christopher G. Ingersoll†, Rebecca A. Dorman†, William G. Brumbaugh†,
Christopher A. Mebane‡ James L. Kunz†, and Doug K. Hardesty†
†U.S. Geological Survey, Columbia Environmental Research Center, Columbia, Missouri
‡U.S. Geological Survey, Boise, Idaho
Table S1. Summary conditions for conducting chronic water-only toxicity tests with White
Sturgeon (Acipenser transmontanus) and Rainbow Trout (Oncorhynchus mykiss) following
standard test methods recommended by the American Society for Testing and Materials
(2013a,b), the U.S. Environmental Protection Agency (2002), and U.S. Gelogical Survey
Columbia Environmental Reserach Center Quality Assurance Project Plan (2010)
Parameter
Conditions
Chemicals
Cadmium chloride, copper sulfate, lead nitrate, zinc chloride
Test type
Flow-through, water-only exposures in intermittent-flow proportional
diluters
Temperature
Sturgeon: 15°C; trout: 12°C
Light
Ambient laboratory light; 16 h light/8 h dark
Test chamber size
9.5 L (20 pieces of smooth stones were placed in the chamber up to swimup life stage during exposure started with newly hatched sturgeon)
Test solution
7L
volume
Water addition
0.25 L/chamber/15 to 30 min (about 2 to 4 volume additions/d)
a
Organisms/chamber C1 and CC: 20 fish (thinned to 10 on test day 25 in sturgeon CC exposure
and on test day 21 in trout CC exposure); C2: 10 fish; C1-R: 30 fish
Loading
<0.5 g fish wet weight/L of solution passing through the chamber in 24 h
and <5 g fish wet weight/L in the chamber at any given time
Replicates
Duration
Age of test
organisms
Feeding
Chamber cleaning
Test water
Dilution series
(nominal
concentrations)
4
Sturgeon: C1: 25 d; C2: 28 d; CC: 53 d; C1-R: 24 d
Trout: C1: 21 d; C2: 28 d; CC: 52 d
Sturgeon: C1: 2 dph; C2: 27 dph; CC: 2 dph; Rep-C1: 1 dph
Trout: C1: 1 dph; C2: 26 dph; CC: 1 dph
Fed brine shrimp nauplii or live aquatic oligochaetes 2 to 3 times daily in
excess (see text for details on timing and amount of food provided
throughout the exposures)
Fish faeces and remaining food siphoned once daily in the morning
Well water diluted with deionized water (see text for details on water
quality characteristics)
Sturgeon:
Cadmium: 0, 0.75, 1.5, 3.0, 6.0, 12 µg/L
Copper: 0, 0.50, 1.0, 2.0, 4.0, 8.0 µg/L
Lead: 0, 5.0, 10, 20, 40, 80 µg/L
Zinc: 0, 25, 50, 100, 200, 400 µg/L
Trout:
Cadmium: 0, 0.75, 1.5, 3.0, 6.0, 12 µg/L
Copper: 0, 10, 20, 40, 80, 160 µg/L
Lead: 0, 10, 20, 40, 80, 160 µg/L
Zinc: 0, 50, 100, 200, 400, 800 µg/L
Aeration
None, except the last week of the sturgeon CC and C2 exposures when
dissolved oxygen decreased to about 5 mg/L
Endpoints
Survival, immobilization, and lack of hiding (newly hatched sturgeon),
total length, dry weight, and biomass
Test acceptability
C1 and C2: ≥80% control survival
criteria
CC: ≥64% control survival
a
C1=chronic life stage 1 exposure, C2=chronic life stage 2 exposure, CC=chronic continuous
exposure, C1-R=repeat C1 exposure.
References:
American Society for Testing and Materials, 2013a. Standard guide for performing early lifesage toxicity tests with fishes, E1241-05. In Annual Book of ASTM Standards, Vol. 11.06,
ASTM International, West Conshohocken, PA, USA.
American Society for Testing and Materials, 2013b. Standard guide for measurement of behavior
during fish toxicity tests, E1711-95 (2008). In Annual Book of ASTM Standards, Vol. 11.06,
ASTM International, West Conshohocken, PA, USA.
U.S. Environmental Protection Agency, 2002. Short-term methods for estimating the chronic
toxicity of effluents and receiving water to freshwater organisms. EPA/821/R-02/013 (5th ed.),
Washington, DC.
U.S. Geological Survey, 2010. USGS-Columbia Quality Assurance Project Plan for an
evaluation of the acute or chronic toxicity of individual chemicals of interest to White Sturgeon
(Acipenser transmontanus) and Rainbow Trout (Oncorhynchus mykiss) in water-only exposures.
U.S. Geological Survey, Columbia Environmental Research Center, Columbia, Missouri.
Figure S1. Examples of hiding behavior and immobilization of White Sturgeon (Acipenser
transmontanus) on test day 8 in the 53-d exposure started with 2-d-post-hatch larvae: fish
actively hiding in a control replicate chamber (A), lack of hiding in the high concentration of
zinc (369 µg Zn/L, B), and immobilization in the high concentration of copper (6.91 µgCu/L, C).