The principal objective of this investigation was to evaluate the effects of mycotoxin contaminated diets (deoxynivalenol (DON)), aflatoxin B1 (AFB1), and fumonisin (FUM)), with or without the use of a commercially available deactivating compound (DC), in young pullets and replacement laying hens on performance, reproductive, serological, and histopathological parameters. In trial 1, experimental treatments consisted of control, low toxin (1 µg DON/g + 1 µg AFB1/g), and high toxin (2 µg DON/g + 2 µg AFB1/g) diets. Pullets fed the high toxin diet had reduced (P<0.05) body weights compared to control and low toxin diets at d 14, 35, 49, 56, and 63. At d 21 and 28, there was a significant interaction observed between mycotoxin and DC inclusion in body weights. Following necropsies (d 35 and 65), relative liver weights and histopathological liver tissue damage were increased (P<0.05) in pullets fed high toxin diets when compared to control and low toxin diets. Relative kidney weights were increased (P<0.05) due to high toxin diet at d 65. Expected negative effects of toxin administration on titer development were not observed. The only interaction observed between mycotoxin administration and DC inclusion in trial 1 was on body weights. In trial 2, experimental treatments consisted of control, DON (9 µg/g) challenge, AFB1 (2 µg/g) + FUM (54 µg/g) challenge, and a mixed challenge (6 µg DON/g, 1 µg AFB1/g, and 27 µg FUM/g). All mycotoxin diets fed to hens negatively influenced (P<0.05) feed efficiency for the trial period spanning weeks 6 through 10 when compared to control diets. Egg production was not affected (P>0.05) by all mycotoxin diets weeks 6 through 10. Relative weights of the liver and kidney were increased (P<0.05) by AFB1+FUM challenge weeks 4 and 9 compared to control diet. The data reported in this study demonstrate that dietary DON and / or AFB1+FUM influence some performance, reproductive, histopathological, and egg quality traits, but by and large, replacement layer pullets seem to be relatively resistant to the mycotoxins evaluated in this trial at the described levels of administration.