These data show the presence/absence and identification of Cryptosporidium species from the results of a molecular survey of various upland river biota aquatic invertebrates, biofilms, mammal droppings and fish guts, gills and faeces. Samples were collected from various upland influenced sites from around Wales between 2012 and 2015 and were collected. Additionally, otter samples from UK-wide project were also tested. Sample collection was primarily undertaken by DURESS researchers at Cardiff University. Sample testing and analysis was performed at the Cryptosporidium Reference Unit, Public Health Wales Microbiology, Swansea. DNA was extracted using a commercially available kit (Gentra PureGene), Qiagen stool and tissue DNA kits for the fish and mammal samples. These data were collected to provide new information required for the production of a catchment pathogen model to inform ecosystems (dis)services analysis of land use change scenarios for the Diversity in Upland Rivers for Ecosystem Service Sustainability (DURESS) project, part of the NERC Biodiversity and Ecosystem Service Sustainability (BESS) BESS Programme. Full details about this dataset can be found at https://doi.org/10.5285/84242834-dc78-49a6-83cb-951edac65d18