This research chose 28 sites to research randomly and properly combined with investigation process as well as means of technologies such as ArcGIS Desktop, ArcGIS Online, ArcGIS Mobile, ArcGIS Collector and Open Data Kit. The result showed that the distribution of fresh water fish diversity in Can Tho city concentrated mainly on canal, farm land, perennial and yearly plant land rural land. The number of species appearing in Can Tho city was divided into 4 levels of biodiversity such as high (accounting for 8,1%), medium (accounting for 82,5%), low (accounting for 2,4%) and very low (accounting for 7%) base on the frequency of fish species in types of land use in Can Tho city.