This is part of Morisatoumi (Connectivity of Hills, Humans and Oceans) project : investigation of 32 rivers. We used environmental DNA for fish diversity survey and also collected environmental data of 32 rivers along the Japan. This is case study with 5 rivers. Those 5 rivers were selected from north to southern part (38.8511 N ~ 31.8417 N), and including mega city and rural area. We estimated the relation with fish diversity and human effect, surrounding, and latitude. The result showed that the diversity reflected the environmental feature. Our study suggested that environmental DNA can be used for biodiversity survey in nation wide range.