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Objectives: Tick-borne diseases have emerged as an increasing medical problem in the world. Being the most prevalent ixodid ticks in Europe, Ixodes ricinus and Dermacentor reticulatus are responsible for transmission of numerous zoonotic pathogens (e.g., human granulocytic anaplasmosis and Lyme borreliosis). Despite their public health significance, studies on the prevalence of tick-borne agents are scare for Eastern Europe. The objective of this study was to examine the prevalence of Anaplasma phagocytophilum, Ehrlichia chaffeensis, and Borrelia burgdorferi sensu lato (B. burgdorferi s. l.) in ixodid ticks from Southeastern Ukraine. Methods: Over a 5-year period (2014–2018), 358 questing and 389 engorged ixodid ticks were collected from Southeastern Ukraine (Zaporizhzhya region). The ticks were identified as Dermacentor marginatus, D. reticulatus, I. ricinus, and Rhipicephalus rossicus. Nucleic acid samples extracted from tick pools were subjected to RT-PCR analyses for A. phagocytophilum, E. chaffeensis, and B. burgdorferi s. l. Results: The examined ixodid ticks tested negative for the aforementioned pathogens with the exception of I. ricinus ticks. For questing I. ricinus ticks, minimum infection rates of A. phagocytophilum and B. burgdorferi s. l. were, respectively, 4.2–7.7% and 8.6–12.7%. Conclusions: These findings will be valuable for medical and veterinary practitioners when risks associated with tick-borne diseases are assessed for southeastern regions of Ukraine.

The Ixodes ricinus tick is the most commonly mentioned transmitter of spirochetes belonging to the Borrelia burgdorferi sensu lato (Bbsl) group. The Bbsl complex currently contains at least 18 genospecies associated with particular reservoir hosts, 8 of which are distributed throughout Europe. The aim of this study was to determine the infestation of Bbsl in ticks Ixodes ricinus and the identification of genospecies of Bbsl group by DNA sequencing. During 2008–2015, ticks Ixodes ricinus removed from humans were collected and tested for the presence of Bbsl. DNA isolation from homogenates was performed by UltraClean BloodSpin DNA kit (MoBio) and by automated instrument Prepito (Perkin-Elmer). RealTime PCR assay based on the specific flagellin sequence amplification for the detection of B. burgdorferi s.l. was performed by EliGene Borrelia UNI kit (Elisabeth Pharmacon, Czech Republic). The kit is used in clinical laboratories in the European Union as an in vitro diagnostic medical device. Internal control was used according to the instruction manual of the kit during isolation of DNA and PCR assays to control process of amplification, and to avoid PCR inhibited samples and false negative results. RealTime positive samples were subsequently DNA sequenced. The annual positivity of the removed ticks ranged from 16 – 33%. The most frequently occurring genospecies was B. afzelii, followed by B. garinii, B. valaisiana, B. spielmanii, and finally, B. burgdorferi sensu stricto. The most important finding is the first identification of B. spielmanii in this region [1]. 1. Bonczek O, Žakovska A, Vargova L, Serý O. Identification of Borrelia burgdorferi genospecies isolated from Ixodes ricinus ticks in the South Moravian region of the Czech Republic. Ann Agric Environ Med. 2015; 22(4): 642–646.

Laimo boreliozė (LB) yra labiausiai paplitusi erkių platinama liga. Ligos sukėlėjus perneša Ixodes rūšies erkės. Didelį borelijų paplitimą lemia platus rezervuarinių šeimininkų ratas. Šį susirgimą gali sukelti kelios B. burgdorferi s. l. komplekso bakterijų rūšys: B. afzelii, B. garinii, B. burgdorferi sensu stricto. Šiame darbe buvo tirtos 134 Ixodes ricinus erkės nuo 8 elninių gyvūnų (4 stirnų ir 4 elnių) ir 73 erkės nuo žolės (elninių teritorijoje). Naudojant dauginės polimerazės grandininės reakcijos (PGR) medodą, 9 - iose (4,3%) iš jų buvo nustatytas užsikrėtimas Borrelia burgdorferi s. l. patogenais, septyniuose mėginiuose buvo identifikuota B. afzelii, viename - B. garinii rūšis. Viename mėginyje buvo nustatyta Borrelia spp., kuri nebuvo identifikuota. Atlikta B. burgdorferi s. l. sekų analizė pagal ospA geną, naudojantis genų banko duomenimis, atskleidė polimorfizmo lygį tarp trijų, žmogui patogeniškų Borrelia rūšių ir parodė padermių pasiskirstymą skirtingose šalyse bei šeimininkuose. Variabiliausios nustatytos B. garinii ospA geno sekos, rasta 361 variabili vieta (nukleotidų įvairovė π=0,072), aptikta 18 sekų variantų. Mažesnis variabilumas nustatytas B. burgdorferi s. s. sekose – aptiktos 266 variabilios vietos (nukleotidų įvairovė π=0,16) ir 12 sekų variantų bei B. afzelii sekose – aptiktos 257 variabilios vietos (nukleotidų įvairovė π=0,17) ir 4 sekų variantai. Didesni skirtumai buvo identifikuoti tarp B. afzelii ospA geno sekų (vidutinis genetinis atstumas... [toliau žr. visą tekstą] Lyme borreliosis (LB) is the most common tick - born disease. The pathogens are transmitted by infected ticks belonging to a species of the genus Ixodes. The high prevalence of Borrelia is determined by a wide circle of reservoir hosts. The disease can be caused by a few bacterial species of B. burgdorferi s.l. complex, such as: B. afzelii, B. garinii and B. burgdorferi sensu stricto. 134 Ixodes ricinus ticks have been explored in the thesis, collected from 8 certine animals (4 roe deer and 4 deer) and 73 ticks collected from grass (in the territory of certine animals). Using multiplex polymerase chain reaction (PCR) method, the infection with Borrelia burgdorferi s. l. pathogens was identified in 9 (4.3%), B. afzelii was identified in seven samples, while B. garinii species was found in one of them. Borrelia spp. was found in one sample, which has not been identified. A sequence analysis B. burgdorferi s. l. implemented according to the ospA gene by using a gene bank data revealed level of polymorphism among three Borrelia species pathogenic to human and showed the distribution of strains in the different countries and hosts. The most variable gene sequences are B. garinii ospA. It is found 361 variable place (nucleotide diversity π=0,072), ant detected 18 sequence variants. Less variability found in B. burgdorferi s. s. sequences: 266 variable places are found and 12 sequence variants (nucleotide diversity π=0,16) and 257 variable places are discovered in B. afzelii... [to full text]