Molecular characterisation of three Ixodes (Pholeoixodes) species (Ixodida, Ixodidae) and the first record of Ixodes (Pholeoixodes) kaiseri from Slovakia

A study of ticks on wildlife was carried out in the area of Levice, Bratislava, Stupava, and Vrbovce (south-western Slovakia) during 2021 and 2022. Overall, 512 ticks were collected from 51 individuals of six wild mammalian species. Eight tick species were identified, namely Dermacentor reticulatus, D. marginatus, Haemaphysalis inermis, H. concinna, Ixodes ricinus, I. hexagonus, and two Ixodes spp. Ixodes hexagonus were collected from northern white-breasted hedgehogs (Erinaceus roumanicus), females belonging to Ixodes spp. were collected from red fox (Vulpes vulpes) and nymphs from European badger (Meles meles). Ixodes hexagonus and the Ixodes spp. were identified morphologically and molecularly based on sequences of fragments of two mitochondrial genes, COI and 16S rRNA. Molecular analysis of Ixodes spp. confirmed the identity of Ixodes kaiseri Arthur, 1957 and I. canisuga (Johnston, 1849). Sequence analyses show that the I. kaiseri isolate from Slovakia is identical to I. kaiseri isolates from Romania, Poland, Germany, Turkey, and Croatia. We demonstrate for the first time the presence of I. kaiseri in Slovakia using both morphological and molecular methods

The population structure of Borrelia lusitaniae Is reflected by a population division of its Ixodes Vector

© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).Populations of vector-borne pathogens are shaped by the distribution and movement of vector and reservoir hosts. To study what impact host and vector association have on tick-borne pathogens, we investigated the population structure of Borrelia lusitaniae using multilocus sequence typing (MLST). Novel sequences were acquired from questing ticks collected in multiple North African and European locations and were supplemented by publicly available sequences at the Borrelia Pubmlst database (accessed on 11 February 2020). Population structure of B. lusitaniae was inferred using clustering and network analyses. Maximum likelihood phylogenies for two molecular tick markers (the mitochondrial 16S rRNA locus and a nuclear locus, Tick-receptor of outer surface protein A, trospA) were used to confirm the morphological species identification of collected ticks. Our results confirmed that B. lusitaniae does indeed form two distinguishable populations: one containing mostly European samples and the other mostly Portuguese and North African samples. Of interest, Portuguese samples clustered largely based on being from north (European) or south (North African) of the river Targus. As two different Ixodes species (i.e., I. ricinus and I. inopinatus) may vector Borrelia in these regions, reference samples were included for I. inopinatus but did not form monophyletic clades in either tree, suggesting some misidentification. Even so, the trospA phylogeny showed a monophyletic clade containing tick samples from Northern Africa and Portugal south of the river Tagus suggesting a population division in Ixodes on this locus. The pattern mirrored the clustering of B. lusitaniae samples, suggesting a potential co-evolution between tick and Borrelia populations that deserve further investigation.This research was financially supported by the Slovak Research and Development Agency (grant number APVV-16-0463), by the Fundação para a Ciência e a Tecnologia by the transitory norm contract DL57/2016/CP1370/CT89 to Ana Cláudia Norte and MARE (MARE-UID/MAR/04292/2020), and by the National Institute of Health Doutor Ricardo Jorge, Lisbon, Portugal. The National Reference Center for Borrelia was supported by the Robert-Koch Institute, Berlin, Germany.info:eu-repo/semantics/publishedVersio