c) Test initiation—The test begins with distribution of newly fertilized eggs or eyed eggs to
each incubation cup. Select number of embryos based on the desired discriminating power for
the test and the number of replicates. At a minimum, apportion 60 embryos per experimental
group among four incubation cups and place two cups in each of two replicate test chambers.
d) Biological observations—Monitor survival by daily inspecting embryos or hatchlings.
Record observations on fish behavior, noting abnormal and normal individuals along with
characteristics of any abnormalities.
During embryo incubation remove dead eggs to prevent fungal infections. Dead eggs can be
distinguished from living eggs by their white color.
When hatching is greater than 95% complete, count post-hatch alevins and release them to
their respective growth chambers. For approximately 2 weeks following hatching, alevins will
feed from their yolk sacs. When the first few alevins begin to swim up, feed fry with brine
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