Pernešėjų platinamų patogenų nustatymas ir genetinė įvairovė smulkiuosiuose graužikuose (Muridae, Cricetidae)

Rodents (Muridae, Cricetidae) represent an important group of mammalian hosts as considered reservoirs for a large number of vector-borne pathogens. However, there still is a lack of knowledge on the geographical distribution, genetic diversity and the reservoir hosts for these microorganisms in Baltic countries. The aim of the study was to investigate the prevalence and genetic diversity of vector-borne pathogens Babesia spp., Rickettsia spp. and Bartonella spp. in various rodent species. A total of 1180 rodents belonged to the eighth species Apodemus flavicollis (n=499), A. agrarius (n=82), Mus musculus (n=12), Micromys minutus (n=77), Myodes glareolus (n=396), Microtus oeconomus (n=69), M. agrestis (n=28) and M. arvalis (n=17) were captured with live or snap traps in 19 locations from coastal and continental areas of Lithuania during 2013–2017. Conventional, semi-nested, nested and real-time PCR were used to amplify different genome regions of Bartonella (ssrA, rpoB, groEL and 16S-23S ITS), Rickettsia (gltA and17kDa) and Babesia (18S rRNA) with subsequent sequence analysis. The overall prevalence of Bartonella pathogens in rodents was 54.8 %, Rickettsia – 28.3%, and Babesia – 2.8%. Small rodents in Lithuania harbor multiple Bartonella species belonging to six genogroups, specifically Bartonella grahamii, B. taylorii, B. tribocorum, B. coopersplainsensis, B. doshiae and B. rochalimae. The highest Bartonella infection rates and the highest species diversity are detected in rodents from coastal areas. B. grahamii and B. taylorii are frequently detected in rodents from both coastal and continental areas of Lithuania, with the high genetic diversity of B. taylorii. The present study represents the first evidence that Babesia pathogens circulating in A. flavicollis, M. minutus, M. glareolus, M. oeconomus and M. agrestis, and the presence of zoonotic Babesia microti ‘Jena/Germany’ strain potentially pathogenic to humans. The high prevalence of Rickettsia helvetica pathogens in A. flavicollis, M. minutus and M. glareolus detected in the present study suggests that these rodents may play an important role as potential reservoir hosts and thus in the maintenance of this pathogen in Lithuania. The findings of our study demonstrated the importance of small rodents as potential reservoirs of zoonotic pathogens in Lithuania.Ekosistemoje graužikai (Muridae, Cricetidae) vaidina svarbų vaidmenį, nes yra svarbūs ektoparazitų šeimininkai ir jų platinamų patogenų rezervuarai. Tačiau Baltijos šalyse vis dar trūksta duomenų apie pernešėjų platinamų patogenų geografinį pasiskirstymą, genetinę įvairovę ir rezervuarus. Darbo tikslas – ištirti pernešėjų platinamų Babesia spp., Bartonella spp. ir Rickettsia spp. patogenų paplitimą ir genetinę įvairovę tarp Lietuvos pakrantėse ir kontinente sugautų graužikų. 2013–2017 m. gyvagaudžiais ir mušamaisiais spąstais 19-oje Lietuvos vietovių sugauta 1180 smulkiųjų graužikų priklausančių aštuonioms rūšims: Apodemus flavicollis (n = 499), A. agrarius (n = 82), Mus musculus (n = 12), Micromys minutus (n = 77), Myodes glareolus (n = 396), Microtus oeconomus (n = 69), M. agrestis (n = 28) ir M. arvalis (n = 17). Bartonella spp. (ssrA, rpoB, groEL ir 16S-23S ITS), Rickettsia spp. (gltA ir 17kDa) ir Babesia spp. (18S rRNR) patogenų nustatymui atlikta standartinė, pusiau-lizdinė, lizdinė ir tikro-laiko PGR. Lietuvoje sugauti smulkieji graužikai yra dažnai infekuoti Bartonella (54,8 %) ir Rickettsia (28,3 %) patogenais, ir retai infekuoti Babesia parazitais (2,8 %). Šešios Bartonella genogrupės – Bartonella grahamii, Bartonella taylorii, Bartonella tribocorum, Bartonella coopersplainsensis, Bartonella doshiae ir Bartonella rochalimae identifikuotos Lietuvos smulkiuosiuose graužikuose. Didžiausias Bartonella spp. užsikrėtimas ir rūšinė įvairovė nustatyta Lietuvos pakrantėse. Lietuvos pakrantėse ir kontinente sugautuose smulkiuosiuose graužikuose dažniausiai nustatytos B. grahamii ir B. taylorii rūšys, su didele B. taylorii genetine įvairove. Microtus oeconomus, Microtus agrestis, Myodes glareolus ir Apodemus flavicollis cirkuliuojantys Babesia pirmuonys yra susiję su žmogui patogeniška Babesia microti ‘Jena/Germany‘paderme. Apodemus flavicollis, Micromys minutus ir Myodes glareolus yra žmogui patogeniškos Rickettsia helvetica platintojai ir vaidina svarbų vaidmenį palaikant šį patogeną Lietuvoje. Lietuvoje smulkieji graužikai yra svarbūs pernešėjų platinamų zoonozinių patogenų rezervuarai.Gamtos mokslų fakultetasBiologijos katedr

Prevalence of Babesia SPP. in Dermacentor Reticulatus and Ixodes Ricinus Ticks in Lithuania

Babeziozė yra infekcinė liga, cirkuliuojanti tarp stuburinių šeimininkų ir vektorių, kurią sukelia skirtingi Babesia pirmuonys. Ixodes ricinus erkės yra laikomos pagrindiniais žmogaus, naminių ir laukinių gyvūnų babeziozės vektoriais. Svarbiausiais šunų babeziozės vektoriais yra laikomos Dermacentor reticulatus erkės. Nors iki šiol Lietuvoje žmogaus babeziozės atvejų dar nebuvo užregistruota, tačiau pastaruoju metu buvo užfiksuota daug šunų babeziozės atvejų. Šio tyrimo tikslas identifikuoti Babesia spp. patogenus ir nustatyti jų paplitimą Dermacentor reticulatus ir Ixodes ricinus erkėse įvairiuose Lietuvos regionuose. Iš viso buvo surinkta 2259 Dermacentor reticulatus ir 370 Ixodes ricinus erkių iš 40 Lietuvos vietovių. Lizdinės PGR metu pagausinta Babesia 18S rRNR geno dalis ir atlikta sekoskaita leido įvertnti Babesia spp. paplitimą Dermacentor reticulatus (1,2 %) ir Ixodes ricinus (9,5 %) erkėse. Babesia spp. paplitimas Dermacentor reticulatus ir Ixodes ricinus erkėse skirtingose vietovėse varijavo nuo 2 % iki 11,4 % ir nuo 3,6 % iki 31,6 %, atitinkamai. Atlikus Dermacentor reticulatus 18S rRNR geno sekų analizę nustatyta, kad 94,1 % (16/17) buvo užsikrėtę Babesia canis ir 5,9 % (1/17) Babesia venatorum. Ixodes ricinus erkėse buvo identifikuotos Babesia microti (9/15; 60 %) ir Babesia venatorum (6/15; 40 %). Dermacentor reticulatus erkėse remiantis dviejų nukleotidų (GA → AG) inversija nustatyti trys Babesia canis genotipai.Babesiosis is an infectious disease circulating worldwide among the vertebrate hosts and vectors which caused by different intraerythrocytic protozoan Babesia. Ixodes ricinus ticks are considered the main vectors of Babesia species causing diseases in human, domestic and wild animals, while other tick Dermacentor reticulatus is recognized as the most important vector of canine babesiosis. In Lithuania up to date, no human cases due to Babesia spp. have been reported, however in the beginning of 21th century were registered many cases of canine babesiosis in dogs. The aim of this study was investigate the presence of Babesia spp. in Dermacentor reticulatus and Ixodes ricinus ticks in various regions in Lithuania, and to assess the risk of Babesia infection in the human population and to better understand epidemiology of canine babesiosis. A total of 2259 Dermacentor reticulatus and 370 Ixodes ricinus were collected from 40 locations in Lithuania. Different regions of the 18S rRNA gene of the genus Babesia were amplified by using nested PCR. The prevalence of pathogens in Dermacentor reticulatus (1.2 %) and Ixodes ricinus (9.5 %) ranged in different locations from 2 % to 11.4 % and from 3.6 % to 31.6 %, respectively. Sequence analyses of DNA from these positive samples indicate the presence of Babesia canis (16/17; 94,1 %) and Babesia venatorum (1/17; 5,9 %) in Dermacentor reticulatus ticks, Babesia microti (9/15; 60 %) and Babesia venatorum (6/15; 40 %) in Ixodes ricinus ticks. Three genotypes of Babesia canis were distinguished on the basis on two nucleotide (GA → AG) substitutions observed in 18S rRNA gene sequences.Vytauto Didžiojo universiteta

Babesia spp. tyrimai Lietuvoje (Investigation of Babesia spp. in Lithuania)

Aplinkotyros katedraVytauto Didžiojo universiteta

Detection of Rickettsia spp. in ticks from different species of migrating passerine birds in Lithuania

eISSN 2300-6706. Abstracts of the XXIV Congress of the Polish Parasitological Society Krakow, 5-8th September 2016, KrakowBirds play an important role in the global dispersal of tick-borne pathogens. During migration, they spread ticks within and between continents. Tick-borne rickettsiae are considered to be emerging, but there is still a lack of data on the occurrence and prevalence of the spotted fever group (SFG) rickettsiae across Europe, especially in the Baltic countries. The aim of this study was to investigate the presence of Rickettsia spp. in ticks from migrating passerine birds. Birds were trapped and examined for the presence of ticks at Ventes Ragas ornithological station, situated in the northwest of Lithuania, during 2013 and 2014. In total, 237 ticks (95 larve and 142 nymphs) were removed from 91 birds of 15 different species. All ticks were identified as Ixodes ricinus. Twenty-three out of 91 infested (25.3%) birds carried Ixodes ricinus ticks infected with Rickettsia spp. Rickettsia spp. DNA was detected in 11.4 % of the tick samples. In total, 6.3% of larvae (6/95) and 14.8% of nymphs (21/142) were positive. Erithacus rubecula was the species of birds with the largest number of infected ticks. This study represents the first investigation of Rickettsia spp. from ticks collected from migrating birdsAplinkotyros katedraBiologijos katedraVytauto Didžiojo universiteta

Molecular characterization of Babesia canis in Dermacentor reticulatus ticks

Aplinkotyros katedraBiologijos katedraGamtos mokslų fakultetasVytauto Didžiojo universiteta

Tick-borne pathogens in ticks collected from domestic dogs and cats in Lithuania

EAN: 9789086862917; e-EAN: 978-90-8686 -837 -7, e-ISBN: 978-90-8686-837 -7. DOI: 10.3920/97 8-90-8686-837-7Biologijos katedraGamtos mokslų fakultetasVytauto Didžiojo universiteta

Babesia microti in rodents from different habitats of Lithuania

Babesia microti (Aconoidasida: Piroplasmida) (Franca, 1910) is an emerging tick-borne parasite with rodents serving as the considered reservoir host. However, the distribution of B. microti in Europe is insufficiently characterized. Based on the sample of 1180 rodents from 19 study sites in Lithuania, the objectives of this study were: (1) to investigate the presence of Babesia parasites in eight species of rodents, (2) to determine the prevalence of Babesia parasites in rodents from different habitats, and (3) to characterize the detected Babesia strains using partial sequencing of the 18S rRNR gene. Babesia DNA was detected in 2.8% rodents. The highest prevalence of Babesia was found in Microtus oeconomus (14.5%) and Microtus agrestis (7.1%) followed by Clethrionomys glareolus (2.3%), Apodemus flavicollis (2.2%) and Micromys minutus (1.3%). In M.minutus, Babesia was identified for the first time. The prevalence of Babesia-infected rodents was higher in the meadow (5.67%) than in the ecotone (1.69%) and forest (0.31%) habitats. The sequence analysis of the partial 18S rRNA gene reveals that Babesia isolates derived from rodents were 99–100% identical to human pathogenic B. microti ‘Jena/Germany’ strai

Investigation of Babesia spp. in sympatric populations of Dermacentor reticulatus and Ixodes ricinus ticks in Lithuania and Latvia

Funding: Research Council of Lithuania (grant number No. MIP-053/2013)The objectives of the present study were to investigate the presence of the Babesia parasites in Dermacentor reticulatus ticks along its current distribution range in Lithuania and Latvia, and Ixodes ricinus in Lithuania, to characterize the detected Babesia spp. using partial sequencing of 18S rRNR gene, and to determine the prevalence of the Babesia pathogens in ticks from different locations of Lithuania and Latvia. From 2013 to 2015, four D. reticulatus nymphs and 2255 questing adults were collected from 40 locations in Lithuania, and 181 questing adult D. reticulatus specimens from 11 locations in Latvia. In Lithuania, 16 questing I. ricinus nymphs and 354 questing adults were collected from 12 locations with the sympatric existence of D. reticulatus and I. ricinus. In Lithuania, Babesia spp. were detected in 1.2% (26/2259) of D. reticulatus and in 9.5% (35/370) of I. ricinus ticks. The overall prevalence of Babesia in D. reticulatus ticks from Latvia was 2.8% (5/181). Sequence analysis of partial 18S rRNA gene of positive samples indicated the presence of B. canis and B. venatorum in D. reticulatus ticks, whereas B. microti and B. venatorum were detected in I. ricinus ticks. Our study is the first investigation on the prevalence and molecular characterization of zoonotic B. canis, B. venatorum, and B. microti in ixodid ticks in Lithuania, and of B. canis in D. reticulatus ticks in Latvia. It is also the first report of B. venatorum in D. reticulatus ticksBiologijos katedraGamtos mokslų fakultetasVytauto Didžiojo universiteta

Occurrence and diversity of vector-borne pathogens in rodents (Muridae, Cricetidae) from Lithuania

Rodents represent an important group of mammalian hosts as considered reservoirs for a large number of vector-borne pathogens. However, there still is a lack of knowledge on the occurrence and diversity of vector-borne pathogens of zoonotic importance in rodents from Baltic countries.The aim of the research was to investigate Bartonella spp., Rickettsia spp. and Babesia spp. prevalence and genetic diversity in various species of rodents captured in Lithuania. Rodents representing eight species – Apodemus flavicollis, Apodemus agrarius, Mus musculus, Micromys minutus, Myodes glareolus, Microtus oeconomus, Microtus agrestis and Microtus arvalis were trapped with live-traps or snap traps in 19 locations of Lithuania during 2013–2016. DNA was extracted from the spleen samples of rodents. Conventional, nested and real-time PCR were used to amplify different genome regions of Bartonella (ssrA, rpoB, groEL and 16S-23S ITS), Rickettsia (gltA and17kDa) and Babesia (18S rRNA) with subsequent sequence analysis. The present study demonstrates that Lithuanian rodents are frequently infected with Bartonella (54.8%), and Rickettsia (28.3%) pathogens and rarely infected with Babesia parasites (2.8%). Sequence analysis of Babesia, Rickettsia and Bartonella isolates showed the presence of Babesia microti, Rickettsia helvetica and multiple Bartonella species belonging to six clades associated with human pathogenic Bartonella grahamii, Bartonella rochalimae and Bartonella tribocorum species, and other species Bartonella taylorii, Bartonella coopersplainsensis and Bartonella doshiae which pathogenicity to humans is still unknown.The findings of our study demonstrated the importance of rodents as considered reservoirs of these vector-borne zoonotic pathogens in LithuaniaAplinkotyros katedraBiologijos katedraGamtos tyrimų centrasVytauto Didžiojo universiteta

Investigation of Babesia spp. infecting Dermacentor reticulatus and Ixodes ricinus ticks in Lithuania

Babesia species, the causative agents of babesiosis, are protozoan blood parasites that are transmitted by ticks to their vertebrate hosts. Ixodes ricinus ticks are considered the main vectors of Babesia species causing diseases in human, domestic and wild animals, while other tick of Ixodidae family Dermacentor reticulatus is recognized as the most important vector of canine babesiosis. In Lithuania up to date, no human cases due to Babesia spp. have been reported, however in the beginning of 21th century were registered many cases of canine babesiosis in dogs. The aim of this study was investigate the presence of Babesia spp. in D. reticulatus and I. ricinus ticks in various regions in Lithuania, and to assess the risk of Babesia infection in the human population and to better understand epidemiology of canine babesiosis. A total of 2259 D. reticulatus and 370 I. ricinus were collected from 40 locations in Lithuania.This study represents the first investigation and characterization of Babesia spp. infecting D. reticulatus and I. ricinus in Lithuania. Different regions of the 18S rRNA gene of the genus Babesia were amplified by using nested PCR. The prevalence of pathogens in D. reticulatus (1,2%) and I. ricinus (9.5%) ranged in different locations from 0% to 6.7% and 0% to 18.5%, respectively. Sequence analyses of DNA from these positive samples indicate the presence of B. canis and B. venatorum in D. reticulatus ticks, B. microti and B. venatorum in I. ricinus ticks. This study represents the first investigation of zoonotic Babesia parasites in LithuaniaAplinkotyros katedraBiologijos katedraGamtos mokslų fakultetasVytauto Didžiojo universiteta