The number of nipples at necropsy was compared against the number of areolas/nipples on PCD 35. Male necropsies were completed on PCD 213. Testes and epididymides from all males were preserved in Bouin fixative and placed in 70% ethanol after 24 h. Histopathologic examination of the testes and epididymides was conducted by Experimental Pathology Laboratories, Inc. (Research Triangle Park, NC), where tissues were sectioned at 4-6 |xm and stained using hematoxylin-eosin.
Necropsy of Matured Female Offspring
Beginning on PCD 214, female offspring of control dams and those treated with 250 mg/kg BW per day were necropsied as described above until all high-dose animals had been examined. Because there was evidence of masculinized anogenital distance on female pups at PCD 25 (PND 3), indicators of masculinization, including anogenital distance, ano-vaginal distance, phallus length (defined here as the distance from the tip of the phallus to the anterior edge of the vaginal orifice), kidney to ovary distance, and vertical kidney to ovary distance were measured.