Rotaviruses, especially the species Rotavirus A (RVA), are worldwide distributed enteric pathogens with high prevalence and pathogenicity in humans and variety of animals. The evidence for frequent intersections between the evolution of human and animal rotaviruses has been well documented. However the research on animal RVA strains is significantly lagging, especially on strains originating in wildlife. Therefore, the main aim of this project is to conduct a synchronized spatiotemporal molecular epidemiology study in order to assess the zoonotic potential of autochthonous RVA in Croatian ecosystem, by following the One Health concept. Our multidisciplinary research idea brings integrity and comprehensiveness in its approach and such concerted effort at the interface of veterinary medicine and public health is considered as warranted. By means of selected methodology, we will have a unique opportunity to get a closer look into genomic structure, genotype constellation, phylogenetic relatedness and zoonotic potential of Croatian autochthonous RVA strains simultaneously detected in domestic and wild animals, humans and environment. Moreover, the NGS technology applied for RVA whole genome sequencing will additionally reveal the diversity of viral metagenomes present in Croatian ecosystem and potentially expose new viral threats of public health importance. This study will be the first one defining a genetic backbone of RVAs in several wildlife species and bring a novel perspective on RVA evolution and the capacity of wildlife RVAs for the emergence into human and domestic animal populations. An autochthonous RVA virus bank will be formed, along with databases enabling future seamless genetic tracking of novel emerging zoonotic strains. This project proposal will install a Reco research group with the aim to boost knowledge, networking and competitiveness of each team member in the European Research Area, resulting in future novel collaborative research initiatives.