Freezing ovarian fluid does not alter how it affects fish sperm swimming performance: creating a cryptic female choice 'spice rack' for use in split-ejaculate experimentation.

Cryptic female choice is often mediated chemically in external fertilizers by ovarian fluid (OF), which can change sperm swimming performance and bias paternity under sperm competition. Assessing cryptic female choice is hindered by the necessity of using fresh gametes and the short time window available to obtain diverse samples from wild animals. Using split-ejaculate experimental designs and samples from lake trout, brown trout and Atlantic salmon, we evaluated whether freezing OF alters the way in which it modifies sperm swimming. Sperm had improved swimming performance in the presence of OF over plain water, and the effect did not depend on whether the OF had previously been frozen. Freezing OF does not seem to alter the way it influences sperm. This allows the researcher to create a 'spice rack' of OF samples that can be used in studies on cryptic female choice, and opens up the possibility to compare animals mating under large spatial and temporal variability.