First detection and subtyping of zoonotic Cryptosporidium in European hedgehogs (Erinaceus europaeus) from Paris region (France) Myriam Thomas1*, Camille Srun2, Bruno Polack1, Pascal Arné2, Mohamed Mammeri1, Aurélie Chevillot1, Karim Tarik Adjou1, Veronica Risco-Castillo1,2 1Anses, INRAE, Ecole Nationale Vétérinaire d'Alfort, UMR BIPAR, Laboratoire de Santé Animale, 94700 Maisons-Alfort, France. 2Ecole Nationale Vétérinaire d'Alfort, Centre Hospitalier Universitaire Vétérinaire de la Faune Sauvage (Chuv-FS), 94700 Maisons-Alfort, France. *myriam.thomas@anses.fr The protozoan parasite Cryptosporidium spp. is responsible of morbidity and mortality, particularly among immunocompromised individuals, children, and elderly people. Given its broad host range encompassing both animals and humans, there is a pressing need to elucidate potential transmission sources. Although European hedgehogs (Erinaceus europaeus) are identified as susceptible hosts, the extent of Cryptosporidium circulation within peri-urban areas in France and the associated public health significance remain uncertain. To address this gap, the presence of Cryptosporidium spp. was investigated from hedgehogs hosted at the Wildlife Hospital from the National Veterinary School of Alfort (CHUV-FS, EnvA). Faecal samples (n = 132) were recovered between June 2019 and February 2020, and analysed for the presence of Cryptosporidium spp. using direct immunofluorescent assays. Positive samples were further characterised by nested PCR targeting the 18S ribosomal RNA (SSU rRNA) and the 60 kDa glycoprotein (gp60) genes, and by sequencing. Additionally, epidemiological data such as sex, age (body weight), origin, cause of admission (nutritional, traumatic, infectious) was compiled and statistically analysed. An overall apparent prevalence of 12.9% of Cryptosporidium spp. was obtained, confirming that European hedgehogs from peri-urban areas of Paris region are reservoirs of Cryptosporidium species with zoonotic potential. Most positive hedgehogs did not show digestive clinical signs. Based on SSU rRNA PCR and sequencing, nine faecal samples were found to be positive for Cryptosporidium parvum, and one for Cryptosporidium erinacei. Nine out of 10 SSU rRNA positive samples were also positive using gp60 PCR, and were assigned to the three following zoonotic subtype families: C. parvum (IIc: n = 7; IId: n = 1) and C. erinacei (XIII: n = 1). To the best of our knowledge, this is the first time of Cryptosporidium description in European hedgehogs in France. Further investigation into the involvement of the European hedgehog in the spread of zoonotic parasites and its implications for public health in France is warranted. Moreover, proactive measures for control are imperative to mitigate transmission among animals and caregivers within Wildlife Centres, as well as among individuals harboring or interacting with these animals.