In this study, we investigated the pathological effects of novel goose parvovirus (NGPV) infection on the skeletal muscle, brain, and intestine of naturally affected ducks suffering from locomotor dysfunction as a new approach for a deeper understanding of this clinical form. For this purpose, a total of 97 diseased ducks, representing 24 flocks of different duck breeds (14–75 days old), were clinically examined. In total, 72 tissue pools of intestine, brain, and skeletal muscle samples were submitted for molecular identification. Typical clinical signs among the examined ducks suggested parvovirus infection. Regarding postmortem examination, all examined ducks showed muscle emaciation (100%) either accompanied by congestion (34%) or paleness (66%). Slight congestion, either in the brain (82.5%) or intestine (75.25%), was predominantly detected. Based on molecular identification, the intestine had the highest percentage of positive detection (91.7%), followed by the skeletal muscle (70.8%), and the brain (20.8%). The main histopathological alterations were myofibre atrophy and degeneration, marked enteritis accompanied by lymphocytic infiltration in the lamina propria and submucosa, while the affected brains showed vasculitis, diffuse gliosis, and Purkinje cell degeneration in the cerebellum. Next-generation sequencing further confirmed the presence of a variant strain of goose parvovirus (vGPV) that is globally known as NGPV and closely related to Chinese NGPV isolates. Using immunohistochemistry, the NGPV antigen was positively detected in the muscle fibres, enterocytes, and Purkinje cells in the cerebellum. These findings provided proof of the involvement of virus replication in the locomotor disorders linked to NGPV infection in ducks.