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 Dermacentorreticulatus, D.marginatus, Haemaphysalisinermis, H.concinna, Ixodesricinus, I.hexagonus, and two Ixodes spp. Ixodeshexagonus were collected from northern white-breasted hedgehogs (Erinaceusroumanicus), females belonging to Ixodes spp. were collected from red fox (Vulpesvulpes) and nymphs from European badger (Melesmeles). Ixodeshexagonus 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 Ixodeskaiseri 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.

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.

Vectors of disease at the northern distribution limit of the genus Dermacentor in Eurasia: D. reticulatus and D. silvarum.

The two ixodid tick species Dermacentor reticulatus (Fabricius) and Dermacentor silvarum Olenev occur at the northern distribution limit of the genus Dermacentor in Eurasia, within the belt of [Formula: see text] latitude. Whilst the distribution area of D. reticulatus extends from the Atlantic coast of Portugal to Western Siberia, that of D. silvarum extends from Western Siberia to the Pacific coast. In Western Siberia, the distribution areas of the two Dermacentor species overlap. Although the two tick species are important vectors of disease, detailed information concerning the entire distribution area, climate adaptation, and proven vector competence is still missing. A dataset was compiled, resulting in 2188 georeferenced D. reticulatus and 522 D. silvarum locations. Up-to-date maps depicting the geographical distribution and climate adaptation of the two Dermacentor species are presented. To investigate the climate adaptation of the two tick species, the georeferenced locations were superimposed on a high-resolution map of the Köppen-Geiger climate classification. The frequency distribution of D. reticulatus under different climates shows two major peaks related to the following climates: warm temperate with precipitation all year round (57%) and boreal with precipitation all year round (40%). The frequency distribution of D. silvarum shows also two major peaks related to boreal climates with precipitation all year round (30%) and boreal winter dry climates (60%). Dermacentor silvarum seems to be rather flexible concerning summer temperatures, which can range from cool to hot. In climates with cool summers D. reticulatus does not occur, it prefers warm and to a lesser extent hot summers. Lists are given in this paper for cases of proven vector competence for various agents of both Dermacentor species. For the first time, the entire distribution areas of D. reticulatus and D. silvarum were mapped using georeferenced data. Their climate adaptations were quantified by Köppen profiles.

Local Population Structure and Seasonal Variability of Borrelia garinii Genotypes in Ixodes ricinus Ticks, Slovakia.

Lyme disease (LD) is the most common tick-borne human disease in Europe, and Borrelia garinii, which is associated with avian reservoirs, is one of the most genetically diverse and widespread human pathogenic genospecies from the B. burgdorferi sensu lato (s.l.) complex. The clinical manifestations of LD are known to vary between regions and depend on the genetic strain even within Borrelia genospecies. It is thus of importance to explore the genetic diversity of such pathogenic borreliae for the wide range of host and ecological contexts. In this study, multilocus sequence typing (MLST) was employed to investigate the local population structure of B. garinii in Ixodes ricinus ticks. The study took place in a natural wetland in Slovakia, temporally encompassing spring and autumn bird migration periods as well as the breeding period of resident birds. In total, we examined 369 and 255 ticks collected from 78 birds and local vegetation, respectively. B. burgdorferi s.l. was detected in 43.4% (160/369) of ticks recovered from birds and in 26.3% (67/255) of questing ticks, respectively. Considering the ticks from bird hosts, the highest prevalence was found for single infections with B. garinii (22.5%). Infection intensity of B. garinii in bird-feeding ticks was significantly higher than that in questing ticks. We identified ten B. garinii sequence types (STs) occurring exclusively in bird-feeding ticks, two STs occurring exclusively in questing ticks, and one ST (ST 244) occurring in both ticks from birds and questing ticks. Four B. garinii STs were detected for the first time herein. With the exception of ST 93, we detected different STs in spring and summer for bird-feeding ticks. Our results are consistent with previous studies of the low geographic structuring of B. garinii genotypes. However, our study reveals some consistency in local ST occurrence and a geographic signal for one of the clonal complexes.

Geographical distribution, climate adaptation and vector competence of the Eurasian hard tick Haemaphysalis concinna.

The ixodid tick Haemaphysalis concinna Koch, 1844 is a proven vector of tick-borne encephalitis (TBE) virus and Francisella tularensis, the causative agent of tularaemia. In the present study, up-to-date maps depicting the geographical distribution and climate adaptation of H. concinna are presented. A dataset was compiled, resulting in 656 georeferenced locations in Eurasia. The distribution of H. concinna ranges from the Spanish Atlantic coast to Kamchatka, Russia, within the belt of 28-64°â€¯N latitude. H. concinna is the second most abundant tick species after Ixodes ricinus collected from birds, and third most abundant tick species flagged from vegetation in Central Europe. To investigate the climate adaptation of H. concinna, the georeferenced locations were superimposed on a high-resolution map of the Köppen-Geiger climate classification. A frequency distribution of the H. concinna occurrence under different climates shows three peaks related to the following climates: warm temperate with precipitation all year round, boreal with precipitation all year round and boreal, winter dry. Almost 87.3 % of all H. concinna locations collected are related to these climates. Thus, H. concinna prefers climates with a warm and moist summer. The remaining tick locations were characterized as cold steppes (6.2%), cold deserts (0.8%), Mediterranean climates (2.7%) or warm temperate climates with dry winter (2.9%). In those latter climates H. concinna occurs only sporadically, provided the microclimate is favourable. Beyond proven vector competence pathogen findings in questing H. concinna are compiled from the literature.

Origins of recently emerged foci of the tick Dermacentor reticulatus in central Europe inferred from molecular markers.

The ornate dog tick Dermacentor reticulatus is vector of several blood parasites, including Babesia canis, a causative agent of babesiosis. The geographical range of D. reticulatus in Europe is discontinuous with a gap separating eastern and western macroregions. New foci observed in several locations in western and central Europe were considered an expansion of the western population, including foci in western Poland. In the present paper we used molecular markers to identify the origins of these foci, and we compared their genetic polymorphism to D. reticulatus collected in sites situated within the eastern population. The overall polymorphism in mt 16S rDNA was low, and all sites from the western population shared the same haplotype suggesting the expansion in this area. In the marker 5.8S rDNA-ITS2 we found no differences in polymorphism between sites from eastern Poland (eastern population), and newly emerged foci in western Poland considered a putative expansion zone of the western population. However, the sites from western Poland differed considerably from nearby German site. Our results show that foci in western Poland could not have originated from D. reticulatus from the western population, as previously thought. We found that the state border following river hinders considerably gene flow between adjacent sites what suggest that natural dispersal of D. reticulatus by wildlife is unlikely, and the emergence of new foci should rather be contributed to human-associated dispersal. We propose that livestock, and pets travelling with their owners are the most probable source of new foci, and they can easily transfer ticks within a country but not between countries.

Emergence of tick-borne pathogens (Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Ricketsia raoultii and Babesia microti) in the Kyiv urban parks, Ukraine.

To date, only limited data about the presence of ticks and circulation of tick-borne pathogens in urban parks of Kyiv in northern Ukraine are available. In total, 767 ticks (696 Ixodes ricinus and 69 Dermacentor reticulatus) collected in seven urban parks and one suburban oak wood park in Kyiv were individually analyzed by the PCR assays. Tick-borne pathogens, namely spirochetes from Borrelia burgdorferi sensu lato complex, Anaplasma phagocytophilum, and Babesia microti, were detected in 11.1% of tested I. ricinus ticks. In total, 4% of I. ricinus ticks tested positive for the presence of B. burdorferi s.l. (Borrelia afzelii and Borrelia garinii), 5.2% for A. phagocytophilum, and Ba. microti was confirmed in 1.9% of examined ticks. Mixed infections were recorded in four DNA samples, representing the prevalence of 0.6%. One female and two I. ricinus nymphs were simultaneously infected with B. afzelii and A. phagocytophilum, and one female carried B. afzelii and Ba. microti. In addition, 10.1% of D. reticulatus ticks tested positive for Rickettsia raoultii. Identification of infectious agents and their diversity, assessment of the relative epidemiological importance and determination of the prevalence in questing ticks from central parts of the cities are crucial steps towards the tick-borne diseases surveillance in urban environment.

Geographical distribution of Dermacentor marginatus and Dermacentor reticulatus in Europe.

The goal of this paper is to present up-to-date maps depicting the geographical distribution of Dermacentor species in Europe based on georeferenced sampling sites. Therefore, a dataset was compiled, resulting in 1286 D. marginatus (Sulzer, 1776) and 1209 D. reticulatus (Fabricius, 1794) locations. Special emphasis is given to the region of the European Alps depicting a presumable climate barrier of the mountains and to overlaps in the distribution of both species as well as on the situation in eastern European countries. For the latter newly described Dermacentor findings comprise 59 locations in Romania and 62 locations in Ukraine. The geographical distributions of both species in Europe range from Portugal to Ukraine (and continue to the east of Kazakhstan). Although it is well known that D. marginatus is adapted to a warmer and drier climate at more southern latitudes and D. reticulatus to a moderately moist climate at more northern latitudes, the distribution limits of both species were not well known. Here, the northern and southern distribution limits for both species in Europe, as determined from the georeferenced database, were specified for D. marginatus by the belt of 33-51° N latitude and for D. reticulatus by the belt of 41-57° N latitude. Thus, overlapping species distributions were found between 41° N and 51° N.