Bufonid toads generally possess cardiotoxic steroids called bufadienolides as defensive chemicals. Although knowledge of the life stages at which the toad species possess the poison is important for our understanding of diversity of toxicity among bufonid toads, this knowledge is limited. In the present study, we revealed that the Japanese common toad, Bufo japonicus formosus, possesses toxins at the unfertilized egg stage by conducting a bioassay experiment. Recent studies documented that hatchlings of B. j. formosus have lethal toxic effects on native frog tadpoles (Rana pirica) in the invasive area of the toad (Hokkaido). In our bioassay experiment using R. pirica tadpole as a predator, no tadpoles died when they did not consume any prey item during two-days experimental period. However, approximately 90% of R. pirica tadpoles immediately died when they consumed an unfertilized egg of B. j. formosus. These results suggest that the toxin at the early life stages of B. j. formosus is, at least partly, provided from female parent.