Detection of canine babesiosis causative agent Babesia canis indetrmacentor reticulatus ticks in Lithuania and Central Europe

Dermacentor reticulatus ticks are considered as an important vector of different viruses, bacteria, protozoa, which causes a variety of diseases in animals and humans. D. reticulatus is the most important vector of Babesia canis protozoan. B. canis, also known as large canine Babesia, can cause severe disease in infected dogs. The most frequent causative agent of canine babesiosis in Central Europe is B. canis canis. Recently, many new endemic foci of this disease have been reported from European countries. Growing incidence of canine babesiosis was recorded also in Lithuania during the last decade. Information on prevalence of babesia pathogens is scarce, especially in Lithuania, where the large-scale investigation of babesia pathogen in D. reticulatus tick never has been done before. The aim of our study was to investigate the prevalence of B. canis in D. reticulatus ticks collected in 34 locations in Lithuania, Poland, Slovakia, Latvia and Ukraine by using molecular methods. A total of 1331 ticks were collected during April – May of 2013, and examined for the presence of Babesia DNA. For pathogen detection, nested PCR with external (BTF1, BTR1) and internal (BTF2, BTR2) primers, which amplify partial region of the 18S rRNA gene of Babesia species, was carried out. In total 0.95 % of all tested samples were positive for B. canis. In Lithuania prevalence of B.canis ranged from 0 to 6.7% in different locations. In Poland, B.canis pathogens were detected with prevalence ranged in different locations from 0% to 2.7%. No positive samples were found in investigated locations of Latvia, Slovakia and Ukraina. Sequence analysis showed that all Babesia isolates from ticks belonged to B. canis canis subspecies. Analysed 18S rRNA gene sequences were 98–100% similar to the sequences of B. c. canis isolated from ticks in other European countriesBiologijos katedraVytauto Didžiojo universiteta

Genetic diversity of Ixodes ricinus and Ixodes persulcatus ticks based on microsatellite analysis

The application of molecular markers to the study of ticks and mites has recently yielded new insights into their population structures and taxonomic relationships. Ticks have been studied at individual, population and species level. At the individual level these range from general approaches, such as AFLP, RAPD or DALP, to highly specific microsatellite analysis. Ixodes tick genera are distributed all over the world. Ixodes ricinus and Ixodes persulcatus ticks are most widespread in Europe and Asia and have significance in medicine, veterinary and economy. The aim of this study was to evaluate I. ricinus and I. persulcatus ticks genetic diversity using microsatellites markersBiologijos katedraVytauto Didžiojo universiteta

Investigation of genetic diversity of Ixodes ricinus ticks based on mitochondrial DNA and microsatellite markers

The application of molecular markers to the study of ticks has recently provided new insights into their population structures and phylogenetic relationships (Navajas and Fenton 2000). Use of microsatellites as genetic markers in population genetic studies has increased rapidly over the last years including several species of Ixodidae (Roed et al. 2006). Mitochondrial DNA (mtDNA) markers have also proved popular for studying of the systematic of ticks and their population structure. The aim of this study was to evaluate genetic diversity of I. ricinus ticks in Baltic countries using mtDNA and microsatellites markers. We screened three mitochondrial gene sequences 16S rRNA, cytb and control region sequences among 35 I.ricinus and ticks, collected from 16 locations: in Lithuania (4 locations), Latvia (4), Estonia (7) and Slovakia (1). Cytb gene was the most conservative gene that had only 4 haplotypes in Baltic countries, with control region sequences was detected 13 haplotypes and the most haplotypes (16 haplotypes) was detected in 16S rRNR gene sequences. We used four microsatellite loci for investigation of genetic diversity among 180 I.ricinus and ticks, collected from 16 locations in Lithuania (6 locations), Latvia (5) and Estonia (5). A total of 79 alleles were detected: the highest number of alleles per locus (25) was observed in samples from Latvia, followed in samples from Lithuania (20), and the lowest in samples from Estonia (18). The number of observed alleles between loci ranged from 5 to 10, and the observed heterozigosities between 0.2 and 0.8. Analysis of molecular variance (AMOVA) showed that genetic diversity of I. ricinus within sampling locations yielded 79%, and among locations 21% of the total genetic diversity. The PCA analysis not revealed distinct clustering for samples from different locations. Samples of different origin showed to a varying extent overlapsBiologijos katedraVytauto Didžiojo universiteta

Prevalence of Rickettsia spp. in Dermacentor reticulatus ticks in Lithuania and in Central Europe

Dermacentor reticulatus (also known as meadow tick) is one of the Ixodidae family ticks, which expanded its distribution range in Europe recently. Such migration of ticks partly related to the changing climate conditions. D. reticulatus is also considered as an important vector of different viruses, bacteria, protozoa, which causes a variety of diseases in animals and humans. One of those examples is obligate intracellular bacteria belonging to Rickettsia spp. genus. In D. reticulatus ticks most prevalent rickettsias species are R. slovaca, R. helvetica, and R. raoultii, among which R. slovaca and R. helvetica are considered as pathogenic species for humans. However, data about the current Rickettsia spp. pathogens spread in Lithuania and Europe is still missing. The aim of this research was to determine the prevalence of Rickettsia spp. in D. reticulatus ticks in Lithuania and Europe. In the present study were collected and tested by molecular methods 1402 ticks in Lithuania (1077 ticks) and other European countries (325 ticks). In Lithuania ticks were collected from 21 different locations: Jonava (10) Užunvėžiai (1), Viešvilė (1), Raudonė (3), Kintai (2), Kalnėnai (17), Jurbarkas (1), Vėžininkai (1), Seredžius (0), Prienai (1), Baištrakis (2), Karmėlava (8), Ukmergė (37), Degučiai (32), Marimontas (33), Ivoniškės (45), Ventė (40), Klaipėda (88), Rusnė (46), Vorusnė (188), and Šakiai district (137). Ticks also were collected from different areas in Slovakia (52), Poland (189), Latvia (14), Ukraine (11), and Croatia (59). All samples were gathered from grass (flag method) and identified using standard morphological identification keys, determining the tick species, sex and development stage. DNA extraction from ticks was performed using 2.5% ammonia solution and infection with pathogens determined by using standard polymerase chain reaction (PCR) and horizontal electrophoresis in agarose gel. [...]Biologijos katedraVytauto Didžiojo universiteta

Genetic diversity of Ixodes ricinus hard tick in Baltic countries based on mtDNA sequencing

Biologijos katedraGamtos mokslų fakultetasVytauto Didžiojo universiteta

Ixodes ricinus erkių genetinė įvairovė, paremta mikrosatelitų žymenimis

Ixodes genties erkės priklauso Ixodes ricinus kompleksui, kurį sudaro 14 rūšių, paplitusių visame pasaulyje, taip pat ir Europoje. I. ricinus erkės parazituoja įvairių rūšių stuburinius gyvūnus ir yra įvairių infekcinių ligų sukėlėjai. Kadangi patogenų paplitimas erkėse ir šeimininkų specifiškumas priklauso nuo erkių genetinės struktūros, svarbu ištirti iksodinių erkių genetinę įvairovę. Šio tyrimo tikslas – įvertinti I. ricinus erkių genetinę įvairovę ir populiacijų genetinę struktūrą Baltijos šalyse panaudojant mikrosatelitų žymenis. Iš viso buvo ištirta 180 erkių (170 I. ricinus ir 10 I. persulcatus), kurios buvo surinktos 18-oje skirtingų Lietuvos, Latvijos ir Estijos vietovių. Šešios populiacijos, po dvi iš kiekvienos šalies, buvo naudotos genetinės įvairovės tyrimams. Atlikus mikrosatelitinių lokusų analizę buvo nustatyta, kad alelių skaičius tirtuose lokusuose svyravo nuo 5 iki 11. Tikėtinas heterozigotiškumas I. ricinus populiacijose svyravo nuo 0,621 iki 0,8, o stebimas heterozigotiškumas svyravo nuo 0,304 iki 0,634, ir tai rodo didelį nuokrypį nuo Hardy-Weinberg pusiausvyros. Didžiausias heterozigotiškumo lygis buvo pajūrio vietovėse, kur didelę įtaką erkių genetinei įvairovei daro migruojantys paukščiaiIxodes ricinus belongs to Ixodes ricinus complex composed of 14 species of ticks distributed worldwide and the most widely spread tick species in Europe I. ricinus is a major ixodid tick, involved in the transmission of a number of diseases to animals and humans in Eurasia. Genetic diversity of these tick populations may have implications on disease transmission. In order to investigate genetic diversity and to characterize the genetic structure of I. ricinus populations in the Baltic countries microsatellite markers described in literature were used. A total of 180 ticks (170 I. ricinus and 10 I. persulcatus) collected from 18 locations in Lithuania, Latvia and Estonia were examined based on four polymorphic microsatellite loci. Genetic diversity of six I. ricinus populations from Lithuania (n = 60), Latvia (n = 60), and Estonia (n = 60) were compared. The number of observed alleles between loci ranged from 5 to 11. Observed heterozygosities were lower than expected at most loci. Mean estimates of expected heterozigosities (He) over loci and populations varied from 0.621 to 0.800, and observed heterozygosity (Ho) from 0.304 to 0.634. F statistics were calculated to analyze the differences between observed and expected heterozygosities, and to obtain genetic distances between populations. Two loci deviated significantly from Hardy–Weinberg equilibrium. However, the highest observed heterozygosity (Ho) was found in coastal locations and lowest – in continental parts, which shows the importance of birds as hosts to tick population structureBiologijos katedraVytauto Didžiojo universiteta

Prevalence of tick-borne pathogens in Dermacentor reticulatus ticks in the northern boundary of the distribution in Central Europe

Biologijos katedraGamtos mokslų fakultetasVytauto Didžiojo universiteta

Prevalence of tick-borne pathogens in Dermacentor reticulatus ticks in Lithuania and in Central Europe

Biologijos katedraGamtos mokslų fakultetasVytauto Didžiojo universiteta

Current distribution of Dermacentor reticulatus ticks in Baltic countries

Biologijos katedraGamtos mokslų fakultetasVytauto Didžiojo universiteta

Occurrence of Dermacentor reticulatus ticks in the northern boundary in Central Europe

Biologijos katedraGamtos mokslų fakultetasVytauto Didžiojo universiteta