Variability of species of Babesia Starcovici, 1893 in three sympatric ticks (Ixodes ricinus, Dermacentor reticulatus and Haemaphysalis concinna) at the edge of Pannonia in the Czech Republic and Slovakia.

The distribution, variability and host specificity of species of Babesia Starcovici, 1893 were studied in questing ticks collected on the northwestern edge of the Pannonian Basin in the south-easternmost part of the Czech Republic and in western Slovakia. The area is characterised by relatively natural floodplain habitats and the sympatric occurrence of three tick species possessing wide host spectra, namely Ixodes ricinus (Linnaeus), Dermacentor reticulatus (Fabricius) and Haemaphysalis concinna Koch. Analysis was carried out on 1,408 I. ricinus, 2,999 D. reticulatus and 150 H. concinna altogether, collected from 59 localities. We documented the presence of Babesia spp. not only in I. ricinus but also in H. concinna in the Czech Republic. Two isolates from I. ricinus were classified as B. venatorum Herwaldt, Cacciò, Gherlinzoni, Aspöck, Slemenda, Piccaluga, Martinelli, Edelhofer, Hollenstein, Poletti, Pampiglione, Löschenberger, Tura et Pieniazek, 2003 (formerly determined as Babesia sp. EU1), which is a zoonotic parasite and can cause human babesiosis. The rest of our amplicons were very similar to B. canis (Piana et Galli-Valerio, 1895), which is usually transmitted by D. reticulatus. Despite the huge amount of examined samples, all D. reticulatus ticks were Babesia-free. Due to this finding, we did not consider our obtained isolates to be B. canis, but other closely related species possessing a similar sequence of the studied portion of 18S rDNA. Although this genetic marker is most frequently used in PCR-based diagnostic methods of babesias, its low variability compromises its reliability in studies based only on this marker.

Diversity of Babesia spp. in cervid ungulates based on the 18S rDNA and cytochrome c oxidase subunit I phylogenies.

Free ranging ungulates, represented in Europe mostly by several deer species, are important hosts for ticks and reservoirs of tick-borne infections. A number of studies have focused on the prevalence of tick borne pathogens in deer chiefly with the aim to determine their potential role as reservoir hosts for important human and livestock pathogens. However, genetic similarity of Babesia spp. forming a group commonly termed as a clade VI that accommodates the deer piroplasms, complicates this task and has led to the description of a bewildering array of poorly characterised strains. This study aims to resolve this issue by using two independent genetic loci, nuclear 18S rRNA and mitochondrial cytochrome c oxidase subunit I genes, used in parallel to identify Babesia isolates in free-ranging red, sika, and roe deer in two areas of their co-occurrence in the Czech Republic. The COX1 loci, in contrast to 18S rRNA gene, shows a clear difference between interspecific and intraspecific variation at the nucleotide level. The findings confirm B. divergens, Babesia sp. EU1 and B. capreoli in studied deer species as well as common presence of another unnamed species that matches a taxon previously referred to as Babesia sp. or Babesia cf. odocoilei or Babesia CH1 group in several other sites throughout Europe. The invasive sika deers enter the life cycle of at least three piroplasmid species detected in native deer fauna. The presence of B. divergens in both sika and red deer in an area where bovine babesiosis is apparently absent raises important questions regarding the epidemiology, host specificity and taxonomic status of the parasite.