Toxicity of Wescodyne(R) and Betadine(R) to eyed eggs was not adversely affected by water hardness (as calcium and magnesium) or by exposure periods up to 60 min. Both iodophors were much more toxic below pH 6.0 than at pH 8.0. In general Wescodyne was slightly more toxic than Betadine. Significant egg loss occurred if freshly fertilized eggs were water‐hardened in either iodophor at 100 ppm of iodine, but egg loss at 25 ppm of iodine or at 100 ppm if the eggs were disinfected 30 min after water hardening was comparable to the control. Also, there was no effect on the egg mortality or fry development following single or multiple exposures after eggs were water hardened. At pH 6.0 and above, Wescodyne and Betadine at 100 ppm iodine in a 15‐min dip would be safe to use on rainbow trout eggs at any stage of development after water hardening. Recommendations and precautions for hatchery use are given.