Broodfish of Atlantic salmonSalmo salarwere fed either thiamin-deficient diets fortified with 0.05% and 0.1% amprolium chloride (Amp, a thiamin antagonist) or thiamin-sufficient diets (0% Amp and ASD2-30, a U.S. Fish and Wildlife Service open-formula feed) for 6 months before spawning. We observed Cayuga syndrome (CS) only in the fry produced from broodfish fed the amprolium diets. The percent fry mortality due to CS was less than 1% in the 0.05% Amp group and was more than 3% in the 0.1% Amp group. Free thiamin was the predominant form of thiamin in the eggs of all broodfish; concentrations were two orders of magnitude greater than that of thiamin pyrophosphate (TPP) and thiamin monophosphate (TMP). Free thiamin was greatest in eggs of broodfish fed the thiamin-sufficient diets. Total thiamin and TMP in red blood cells (RBCs) of broodfish fed the thiamin-deficient diets were reduced to the initial levels by the end of the study. Levels of free thiamin in RBCs were similar in all treatment groups at spawning, as were levels of TPP. Amprolium feeding significantly reduced total thiamin levels in heart, liver, and muscle but not in kidney. Except for thiamin found in eggs, TPP was the predominant form of thiamin in all the other tissues, whereas free thiamin was the least predominant. The data support the hypothesis that thiamin deficiency may cause Cayuga syndrome in Atlantic salmon sac fry. The low incidence of the syndrome in this study may be due to the short duration of the feeding trial or to the low levels of amprolium added to the diets.