Red Foxes (Vulpes vulpes) and European Badgers (Meles meles) as Overlooked Wildlife Hosts of Canine Parvovirus in Slovakia: First Evidence by Molecular Characterization and Virus Isolation

Wild carnivores are increasingly recognized as hosts or reservoirs of canine parvovirus (CPV), a major pathogen of dogs. To investigate CPV circulation in Central Europe, we examined 221 red foxes (<i>Vulpes vulpes</i>) and 53 European badgers (<i>Meles meles</i>) from seven Slovakian regions. Small intestines (<i>n</i> = 86), rectal swabs (<i>n</i> = 123), and feces (<i>n</i> = 65) were tested by real-time PCR, and positives were sequenced for the <i>VP2</i> gene. Virus isolation was performed on MDCK cells. CPV was detected in 10.9% (30/274) of samples, with a prevalence of 10.9% (24/221) in foxes and 11.3% (6/53) in badgers. Phylogenetic analysis revealed co-circulation of CPV-2a, CPV-2b, and CPV-2c, with CPV-2b being prevalent (20/30, 66.7%) and CPV-2c detected in a single sample (1/30, 3.3%). CPV-2a/2b isolates clustered with European strains, while CPV-2c grouped within the "Asian" lineage. Several sequences carried "Asian-like" signatures (5G, 267Y, 324I, 370R), suggesting transboundary introduction and spillover into wildlife. Two infrequent substitutions were identified: S552I in CPV-2b and I447M in CPV-2a. Viable virus was isolated from all positives, with characteristic CPV-cytopathic effects. This study provides the first molecular and virological evidence of CPV in Central European wildlife. Our findings expand our understanding of CPV diversity in Europe, and underscore wildlife as an integral component of European ecology.