The distribution and prevalence of zoonotic pathogens infecting ixodid ticks in 
Western Europe have been extensively examined. However, data on ticks and 
tick-borne pathogens in Eastern Europe, particularly Ukraine are scarce. The 
objective of the current study was, therefore, to investigate the prevalence of 
Anaplasma phagocytophilum, Anaplasmataceae, Rickettsia spp., Babesia spp., 
Bartonella spp., and Borrelia burgdorferi sensu lato in engorged and questing 
ixodid ticks collected from five administrative regions (oblasts) of Ukraine, 
namely Chernivtsi, Khmelnytskyi, Kyiv, Ternopil, and Vinnytsia. The ticks were 
collected from both wild and domestic animals and from vegetation. Of 524 ixodid 
ticks collected, 3, 99, and 422 ticks were identified as Ixodes hexagonus, 
Ixodes ricinus, and Dermacentor reticulatus, respectively. DNA samples 
individually extracted from 168 questing and 354 engorged adult ticks were 
subjected to pathogen-specific PCR analyses. The mean prevalence in I. ricinus 
and D. reticulatus were, respectively: 10 % (10/97) and 3 % (12/422) for A. 
phagocytophilum; 69 % (67/97) and 52 % (220/422) for members of the 
Anaplasmataceae family; 25 % (24/97) and 28 % (117/422) for Rickettsia spp.; 3 % 
(3/97) and 1 % (6/422) for Babesia spp.; and 9 % (9/97) and 5 % (20/422) for 
Bartonella spp. Overall, between the five cities, there was no significant 
difference in the prevalence of any of the pathogens for the respective ticks (p 
> 0.05). The prevalence of B. burgdorferi s. l. in the questing and engorged I. 
ricinus varied from 0 to 27 % and 14-44%, respectively, with no statistical 
significance identified between the five cities (p > 0.05). In addition to 
reporting the updated data for Kyiv and Ternopil, this study is the first to 
provide the prevalences of the tick-borne pathogens for Chernivtsi, 
Khmelnytskyi, and Vinnytsia. This investigation is also the first to detect 
Neoehrlichia mikurensis in ixodid ticks from Ukraine. These new data will be 
useful for medical and veterinary practitioners as well as public health 
officials when diagnosing infections and when implementing measures to combat 
tick-borne diseases in Ukraine.