Test initiation—See Section 8910B.5b for egg collection. The test begins with
distribution of newly fertilized eggs to each embryo cup. Select a number of embryos based upon
the desired discriminating power for the test and the number of replicates. At a minimum,
distribute 60 embryos per experimental group among four incubation cups and place two cups in
each of two replicate test chambers. Sheepshead minnows hatch in about 7 d at 25°C.6,7 Release
larvae from hatching cup to the test chambers and start feeding. Initially feed a combination of
live brine shrimp nauplii, then shift to brine shrimp and commercial food after 7 to 10 d.6
c) Biological observations—See Section 8910C.2b1)d).
d) Chemical data recording—In addition to those water quality factors described in Section
8910C.2b1)e), measure dilution water salinity at least weekly.
e) Verification of exposure concentrations—See Section 8910C.2a3).
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