Chagas disease is endemic in Latin America where the vector, hematophagous triatomine, is widely distributed. The exposure of women to T. cruzi may result in birth defects. However, congenital transmission of Trypanosoma cruzi from infected mothers to fetuses is scantily studied. Herein, we have developed a guinea pig model of congenital transmission of T. cruzi and evaluated the pathology in fetuses born to mothers that were exposed to T. cruzi before or during gestation. We also show high virulence of a T. cruzi DTU I strain and its capacity to invade fetal tissues even at a very low dose.