<i>Uncinaria stenocephala</i> (northern hookworm) is the major endoparasite in dogs from private dog shelters in the Balkans: presence of benzimidazole susceptible isotype-1 β-tubulin alleles

Zoonotic parasites associated with domestic dogs have been well-studied in the majority of Europe. In the Balkan region, however, there is minimal knowledge of the parasites in dogs in shelters for rehoming in other European countries. This study aimed to investigate parasitic infections in dogs from two private shelters in Pristina, Kosovo. Faecal samples were collected, representing both adult dogs (72%) and puppies (28%). Coproscopic analysis revealed that 88% of dogs were infected with at least one parasite, with hookworms being the most common. Amplicon metabarcoding targeting internal transcribed spacer (ITS)-2 rRNA gene confirmed the presence of only <i>Uncinaria stenocephala</i> in 68% of samples apparently susceptible to benzimidazoles. The canonical F167Y and Q134H isotype-1 β-tubulin of <i>U. stenocephala</i> mutations conferring benzimidazole resistance were not detected. No evidence of <i>Ancylostoma caninum</i> was detected. Molecular analysis confirmed <i>Giardia duodenalis</i> in 18% of samples, with assemblages B, D and C detected. Other parasites detected included <i>Cystoisospora</i> spp. (18%), <i>Toxocara canis</i> (4%), <i>Toxascaris leonina</i> (6%), <i>Trichuris vulpis</i> (32%), <i>Eucoleus aerophilus</i> (10%) and <i>Dipylidium caninum</i> (2%). Co-infections were identified in 48% of the samples. These findings demonstrate a high frequency of gastrointestinal parasites in shelter dogs. The presence of <i>U. stenocephala</i> and <i>T. vulpis</i> points to the challenges with monitoring and managing these parasitic infections in such settings, as these are likely translocated with the rehomed dogs. The frequency of detection of hookworms emphasizes the need for further research into the distribution of hookworms in Europe because of the emerging benzimidazole resistance on other continents.