Schistosoma japonicum, a parasite of significant public health importance in parts of China and Southeast Asia, is a true generalist pathogen with over 40 species of mammals suspected as definitive host reservoirs. In order to characterize levels of parasite gene flow across host species and identify the most important zoonotic reservoirs, S. japonicum larvae (miracidia) were sampled from a range of definitive host species in two contrasting habitat types within Anhui Province, China: a low-lying marshland region, and a hilly region, where animal reservoir populations may be predicted to differ substantially. Miracidia samples were genotyped using seven multiplexed microsatellite markers. Hierarchical F-statistics and clustering analyses revealed substantial geographical structuring of S. japonicum populations within Anhui, with strong parasite genetic differentiation between habitat types. Within most villages, there was very little or no parasite genetic differentiation among host species, suggesting frequent S. japonicum gene flow, and thus also transmission, across species. Moreover, the data provide novel molecular evidence that rodents and dogs are potentially very important infection reservoirs in hilly regions, in contrast to bovines in the marshland regions. The parasite genetic differentiation between habitat types might therefore be associated with contrasting host reservoirs. The high levels of parasite gene flow observed across host species in sympatric areas have important implications for S. japonicum control, particularly in hilly regions where control of infection among wild rodent populations could be challenging.