Interspecific hybridization between two species of Japanese toads, Bufo torrenticola (Bt) and Bufo japonicus formosus (Bjf), was not thought to have occurred due to the differences in their spawning habitats, although their hybridization was reported by previous research in a limited area. Such hybridization could have pivotal effects, such as genetic admixture, on Bt, which has a limited distribution and habitat. In this study, to clarify the details of hybridization, a total of 192 individuals were collected from multiple parapatric regions and an only Bjf allopatric region and analyzed based on morphology, mtDNA sequences, and nuclear DNA microsatellite loci. Each individual was identified based on morphological type and mtDNA lineage, and two mismatched combinations, Bt or an intermediate morphological type and the Bjf mtDNA lineage, were detected. In particular, the combination of Bt type and Bjf lineage was dominant in parapatric regions, and it was considered that asymmetric hybridization in which Bjf became a matriline had occurred. In addition, a Bayesian clustering method based on the microsatellite loci was conducted to detect a genetic admixture of the two toad species. Individuals representing a combination of the Bt type and Bjf lineage in the parapatric regions were largely assigned to the Bt cluster or equally to both clusters. These results indicate that backcross with Bt has been repeated, and therefore the traits of Bt might be maintained in this study.