TDK conference 2016

Research focusing on ticks and tick-borne pathogens has increased during the past decades (both in Hungary and in a worldwide context), but there are still many eco-epidemiological aspects of tick-borne infections that remained hitherto unevaluated. The initial and primary aim of our work was to provide new information on the epidemiology of piroplasmoses in Hungary. Therefore, during the first part of our study, EDTA blood samples were obtained (in October and November, 2015) from 32 horses kept in an area (in Érd), where accumulated cases of canine babesiosis have recently been reported. In addition, EDTA blood samples were collected (in April, 2016) from 100 racka sheep, which are grazing pastures shared with Babesia caballi- and Theileria equi-infected horses (in Hortobágy). The DNA was extracted from these blood samples, and screened for the presence of piroplasm DNA with conventional PCR and sequencing. All horse and sheep blood samples were negative for both "type piroplasms" and unusual piroplasms already reported from these hosts (i.e. in horses: T. equi, B. caballi and B. canis; in sheep: B. motasi, B. ovis and T. equi).

In the second part of our study, samples were collected from wild living carnivores in Northern and Eastern Hungary (i.e. two faecal droppings from wolves, and blood/spleen samples from a wild cat, a least weasel, two beech martens and a badger, as well as ticks from the latter). After DNA extraction these samples were analysed for the presence of piroplasms. Ticks from the badger were morphologically identified as females of Ixodes rugicollis. Based on its cytochrome oxidase subunit I (COI) and 16S rRNA sequences, I. rugicollis phylogenetically clustered together with I. lividus and I. arboricola, i.e. other members of the subgenus Pholeoixodes. In the blood of the badger and in one female I. rugicollis, the DNA of a new Babesia genotype was present, which differed from a piroplasm detected in badgers in Spain, and clustered phylogenetically in the B. microti clade. The blood sample of the badger was also evaluated for tick-borne bacteria in the family Anaplasmataceae, and was found to contain the DNA of Candidatus Neoehrlichia sp. (FU98). This novel genotype has recently been identified in red fox in Austria and the Czech Republic, and is most closely related to Ca. N. lotoris (from raccoons in North America), and has lower sequence identity with the I. ricinus-transmitted zoonotic agent, Ca. N. mikurensis found in Eurasia.

In conclusion, phylogenetic analysis of I. rugicollis (based on two genetic markers) confirms its status in subgenus Pholeoixodes. Ca. Neoehrlichia sp. (FU98) was identified for the first time in badgers and in Hungary. Additionally, a molecularly previously not yet characterized Babesia genotype occurs in badgers in Central Europe.