Ornithological and molecular evidence of a reproducing Hyalomma rufipes population under continental climate in Europe

Background Reports on adult Hyalomma ticks in certain regions of the Carpathian Basin date back to the 19th century. These ticks were thought to emerge from nymphs dropping from birds, then molting to adults. Although the role of migratory birds in carrying ticks of this genus is known from all parts of Europe, in most countries no contemporaneous multiregional surveillance of bird-associated ticks was reported which could allow the recognition of hotspots in this context. Methods Ixodid ticks were collected from birds at seven ringing stations in Hungary, including both the spring and autumn migration period in 2022. Ixodes and Haemaphysalis species were identified morphologically, whereas Hyalomma species molecularly. Results From 38 passeriform bird species 957 ixodid ticks were collected. The majority of developmental stages were nymphs (n=588), but 353 larvae and 16 females were also present. On most birds (n=381) only a single tick was found and the maximum number of ticks removed from the same bird was 30. Tick species were identified as Ixodes ricinus (n=598), Ixodes frontalis (n=18), Ixodes lividus (n=6), Haemaphysalis concinna (n=322), and D. reticulatus (n=1). All twelve Hyalomma sp. ticks (11 engorged nymphs and an unengorged larva) were identified as Hyalomma rufipes based on three mitochondrial markers. This species was only found in the Transdanubian region and along its southeastern border. The Common Blackbird ( Turdus merula ) and the European Robin ( Erithacus rubecula ) were the two main hosts of I. ricinus and I. frontalis , whereas H. concinna was almost exclusively collected form long-distance migrants. The predominant hosts of H. rufipes were reed-associated bird species, the Sedge Warbler ( Acrocephalus schoenobaenus ) and the Bearded Reedling ( Panurus biarmicus ), both harboring these ticks at the end of June (i.e., the nesting period) in southwestern Hungary. Conclusions This study provides ornithological explanation for the regional, century-long presence of adult Hyalomma ticks under continental climate in the Transdanubian Region of the Carpathian Basin. More importantly, the autochthonous occurrence of a H. rufipes population was revealed for the first time in Europe, based on the following observations: (1) the bird species infested with H. rufipes are not known to migrate during their nesting period; (2) one larva was not yet engorged; (3) the larva and the nymphs must have belonged to different local generations; and (4) all H. rufipes found in the relevant location were identical in their haplotypes based on three maternally inherited mitochondrial markers, probably reflecting founder effect. This study also demonstrated that the species of ticks carried by birds were significantly different between collection sites even within a geographically short distance (200 km). Therefore, within a country multiregional monitoring is inevitable to assess the overall epidemiological significance of migratory birds in importing exotic ticks, and also in maintaining newly established tick species. ### Competing Interest Statement The authors have declared no competing interest. * cox 1 : cytochrome c oxidase subunit I