AbstractDichloroacetic acid (DCA) is a by‐product of the chlorine disinfection of water and may occur in treated water at levels exceeding 100 μg/L. Previous studies revealed teratogenic effects, particularly heart malformations, at high doses (900–2,400 mg/kg given on days 6–15 of pregnancy). In a series of three studies, groups of 7–10 Long‐Evans rats were dosed with 1,900 mg/kg of DCA on days 6–8, 9–11, or 12–15; with 2,400 mg/kg on days 10, 11, 12, or 13; and with 3,500 mg/kg on days 9, 10, 11, 12, or 13, in an attempt to determine the most sensitive period and further characterize the heart defect. In a fourth study, six dams were treated with 1,900 mg/kg of DCA days 6–15 of pregnancy, and 56 fetuses were harvested for light microscopy of the heart. Eight control fetuses from four litters were also examined. No heart malformations were seen in the groups treated with 1,900 mg/kg DCA days 6–8 but were present in the group treated on days 9–11 and 12–15, with the higher incidence occurring on days 12–15. Single doses of 2,400 mg/kg DCA given on days 10, 11, 12, or 13 resulted in a much lower incidence of cardiac malformations, which occurred only on days 10 and 12. The high dose of DCA (3,500 mg/kg) did not increase the incidence of heart defects but showed that dosing on day 9 as well as on days 10 and 12 would produce the defect. The defects seen were characterized as high interventricular septal defects (H‐IVSD). Light microscopy showed that the defect was caudal to the semilunar valves, with the anterior right wall of the aorta communicating with the right ventricle. Another aspect of the defect is at the level of the semilunar valves, with the right cusp or sinus of Valsalva in communication with the right ventricle. The defects are discussed more fully and methods for further study sugge