Canine hookworms represent some of the most globally prevalent parasitic nematodes affecting dogs and pose a significant zoonotic risk to humans, in whom they can induce cutaneous larva migrans. Infection with these parasites may lead to blood loss, anaemia, and, in severe cases, mortality—particularly in young puppies. The present study reports a confirmed case of Ancylostoma caninum infection in a 15-month-old dog in the Slovak Republic. The infected dog presented with severe, persistent diarrhoea, and haematological examination revealed a decrease in erythrocyte and haemoglobin levels, as well as mild eosinophilia. Coprological examination confirmed the presence of nematodes belonging to the family of Ancylostomatidae. Following the initiation of anthelmintic therapy, adult individuals were recovered from the faeces. Based on distinct morphological characteristics, the parasites were presumptively attributed to the species A. caninum. Subsequent molecular analysis of the mitochondrial cytochrome c oxidase subunit 1 gene (COX1) and the ribosomal ITS regions definitively confirmed the species A. caninum. Our findings confirm that this is the first molecularly confirmed case of this species in Central Europe. This hookworm is predominantly found in warm and humid climatic regions. Its recent detection in Slovakia, a country characterised by a temperate climate, may suggest a northward expansion of its geographic range, potentially facilitated by ongoing climatic shifts associated with global climate change.