The Prevalence of Tick-Borne Encephalitis Virus in Wild Rodents Captured in Tick-Borne Encephalitis Foci in Highly Endemic Lithuania.

Wild rodents are considered to be one of the most important TBEV-amplifying reservoir hosts; therefore, they may be suitable for foci detection studies. To investigate the effectiveness of viral RNA detection in wild rodents for suspected TBEV foci confirmation, we trapped small rodents (n = 139) in various locations in Lithuania where TBEV was previously detected in questing ticks. Murine neuroblastoma Neuro-2a cells were inoculated with each rodent sample to maximize the chances of detecting viral RNA in rodent samples. TBEV RNA was detected in 74.8% (CI 95% 66.7-81.1) of the brain and/or internal organ mix suspensions, and the prevalence rate increased significantly following sample cultivation in Neuro-2a cells. Moreover, a strong correlation (r = 0.88; p < 0.05) was found between the average monthly air temperature of rodent trapping and the TBEV RNA prevalence rate in cell culture isolates of rodent suspensions, which were PCR-negative before cultivation in cell culture. This study shows that wild rodents are suitable sentinel animals to confirm TBEV foci. In addition, the study results demonstrate that sample cultivation in cell culture is a highly efficient method for increasing TBEV viral load to detectable quantities.

Phylogenetic characterisation of tick-borne encephalitis virus from Lithuania.

The Baltic states are the region in Europe where tick-borne encephalitis (TBE) is most endemic. The highest notification rate of TBE cases is reported in Lithuania, where the incidence of TBE has significantly increased since 1992. A recent study reported 0.4% prevalence of TBE virus (TBEV) in the two most common tick species distributed in Lithuania, Ixodes ricinus and Dermacentor reticulatus, with the existence of endemic foci confirmed in seven out of Lithuania's ten counties. However, until now, no comprehensive data on molecular characterisation and phylogenetic analysis have been available for the circulating TBEV strains. The aim of this study was to analyse TBEV strains derived from I. ricinus and D. reticulatus ticks collected from Lithuania and provide a genotypic characterisation of viruses based on sequence analysis of partial E protein and NS3 genes. The 54 nucleotide sequences obtained were compared with 81 TBEV strains selected from the NCBI database. Phylogenetic analysis of the partial E and NS3 gene sequences derived from 34 Lithuanian TBEV isolates revealed that these were specific to Lithuania, and all belonged to the European subtype, with a maximum identity to the Neudoerfl reference strain (GenBank accession no. U27495) of 98.7% and 97.4%, respectively. The TBEV strains showed significant regional genetic diversity. The detected TBEV genotypes were not specific to the tick species. However, genetic differences were observed between strains from different locations, while strains from the same location showed a high similarity.

Prevalence and Diversity of Haemotropic Mycoplasma Species in Cats and Their Ectoparasites (Fleas and Ticks).

Mycoplasma spp. pathogens frequently cause chronic and acute diseases in cats. The aim of the present study was to investigate the presence and genetic diversity of Mycoplasma spp. in cats and their ectoparasites using PCR and sequence analysis of the 16S rRNA gene. Blood samples were collected from 541 domestic and stray cats in Lithuania. Ectoparasites (153 fleas and 321 ticks) were collected from owned domestic cats that live both outdoors and indoors. Mycoplasma spp. were detected in 7.2% of cat blood samples and 4.4% of Ctenocephalides felis fleas. The sequence analysis revealed the presence of Mycoplasma haemofelis in 1.1% of cats and 'Candidatus Mycoplasma haematominutum' in 4.8% of cats. Ct. felis fleas harboured M. haemofelis. To the best of the authors' knowledge, this is the first report on the prevalence and molecular characterisation of Mycoplasma bacteria in cats in Lithuania and cat fleas in the Baltic States.

Anaplasma phagocytophilum in European bison (Bison bonasus) and their ticks from Lithuania and Poland.

The increasing population of European bison (Bison bonasus) can contribute to the prevalence of zoonotic pathogens. The aim of the present study was to assess the presence of A. phagocytophilum infection in European bison tissues as well as ticks removed from European bison in Lithuania and Poland. A further objective of this work was to compare the detected A. phagocytophilum strains. A total of 85 tissue samples (spleen) of European bison and 560 ticks belonging to two species, Ixodes ricinus (n = 408) and Dermacentor reticulatus (n = 152) were tested. DNA of A. phagocytophilum was detected based on RT-PCR in 40% of the European bison samples, 8.8% of the I. ricinus and 5.9% of the D. reticulatus ticks. Analysis of the obtained partial 16S rRNA gene sequences of A. phagocytophilum revealed the presence of three variants with two polymorphic sites. Furthermore, phylogenetic analysis with partial msp4 gene sequences grouped A. phagocytophilum variants into three clusters. This study revealed that the groEL gene sequences of A. phagocytophilum from European bison and their ticks grouped into ecotype I and only one sequence from Lithuanian European bison belonged to ecotype II. The results of the present study indicated that European bison may play a role as a natural reservoir of A. phagocytophilum.

Molecular Identification and Phylogenetic Analysis of Laelapidae Mites (Acari: Mesostigmata).

The family Laelapidae (Dermanyssoidea) is morphologically and ecologically the most diverse group of Mesostigmata mites. Although molecular genetic data are widely used in taxonomic identification and phylogenetic analysis, most classifications in Mesostigmata mites are based solely on morphological characteristics. In the present study, eight species of mites from the Laelapidae (Dermanyssoidea) family collected from different species of small rodents in Lithuania, Norway, Slovakia, and the Czech Republic were molecularly characterized using the nuclear (28S ribosomal RNA) and mitochondrial (cytochrome oxidase subunit I gene) markers. Obtained molecular data from 113 specimens of mites were used to discriminate between species and investigate the phylogenetic relationships and genetic diversity among Laelapidae mites from six genera. This study provides new molecular data on Laelaps agilis, Laelaps hilaris, Laelaps jettmari, Haemogamasus nidi, Eulaelaps stabularis, Hyperlaelaps microti, Myonyssus gigas, and Hirstionyssus sp. mites collected from different rodent hosts and geographical regions in Europe.

Bartonella spp. detection in laelapid (Mesostigmata: Laelapidae) mites collected from small rodents in Lithuania.

The genus Bartonella contains facultative Gram-negative intracellular bacteria from the family Bartonellaceae that can cause diseases in humans and animals. Various Bartonella species have been detected in rodents' ectoparasites, such as fleas, ticks, mites, and lice. However, the role of laelapid mites (Mesostigmata: Laelapidae) as carriers of Bartonella spp. needs to be confirmed. We aimed to investigate the presence of Bartonella spp. in laelapid mites collected from small rodents in Lithuania using real-time PCR targeting the transfer-messenger RNA/tmRNA (ssrA) gene and to characterize Bartonella strains using nested PCR and sequence analysis of the 16S-23S rRNA intergenic transcribed spacer region (ITS). A total of 271 laelapid mites of five species (Laelaps agilis, Haemogamasus nidi, Eulaelaps stabularis, Myonyssus gigas, and Hyperlaelaps microti) were collected from five rodent species (Apodemus flavicollis, Apodemus agrarius, Clethrionomys glareolus, Micromys minutus, and Microtus oeconomus) during 2015-2016. Bartonella DNA was detected in three mite species L. agilis, M. gigas, and Hg. nidi with an overall prevalence of 11.4%. Sequence analysis of the 16S-23S rRNA ITS region revealed the presence of Bartonella taylorii in L. agilis, Hg. nidi, and M. gigas, and Bartonella grahamii in L. agilis. Our results suggest that laelapid mites are involved in the maintenance of rodent-associated Bartonella spp. in nature. To the best of the authors' knowledge, this is the first study to demonstrate the presence of Bartonella spp. DNA in laelapid mites from small rodents.

Genetic Diversity of Babesia canis Strains in Dogs in Lithuania.

Canine babesiosis is an emerging and rapidly expanding tick-borne disease in central and northeast Europe. In the last two decades, the endemic area of Babesia canis has expanded from central Europe to the Baltic region. This study aimed to investigate the genetic diversity of B. canis strains isolated from naturally infected dogs in different regions of Lithuania using PCR-RFLP and sequence analyses based on a partial region of 18S rRNA and Bc28.1 genes. Blood samples from 149 dogs suspected of having babesiosis were collected in Lithuania during 2016-2017. Based on PCR-RFLP profiles and two nucleotide substitutions observed in 18S rRNA gene sequences, three B. canis genotypes were identified in Lithuania-18S rRNA-A, 18S rRNA-B and 18S rRNA-A/B-with the A/B genotype predominating (83.9%). Based on the obtained PCR-RFLP profiles of the Bc28.1 gene, four B. canis genotypes were identified: Bc28.1-B (53.8%), Bc28.1-34 (20.8%), Bc28.1-A (17.9%), and Bc28.1-34/A or B (7.5%). Sequence analysis of the partial Bc28.1 gene revealed eighteen polymorphic sites and thirteen sequence variants among the Lithuanian samples. The B. canis genotypes obtained were detected with varying prevalences in different regions of Lithuania.

Babesiosis in Southeastern, Central and Northeastern Europe: An Emerging and Re-Emerging Tick-Borne Disease of Humans and Animals.

There is now considerable evidence that in Europe, babesiosis is an emerging infectious disease, with some of the causative species spreading as a consequence of the increasing range of their tick vector hosts. In this review, we summarize both the historic records and recent findings on the occurrence and incidence of babesiosis in 20 European countries located in southeastern Europe (Bosnia and Herzegovina, Croatia, and Serbia), central Europe (Austria, the Czech Republic, Germany, Hungary, Luxembourg, Poland, Slovakia, Slovenia, and Switzerland), and northern and northeastern Europe (Lithuania, Latvia, Estonia, Iceland, Denmark, Finland, Sweden, and Norway), identified in humans and selected species of domesticated animals (cats, dogs, horses, and cattle). Recorded cases of human babesiosis are still rare, but their number is expected to rise in the coming years. This is because of the widespread and longer seasonal activity of Ixodes ricinus as a result of climate change and because of the more extensive use of better molecular diagnostic methods. Bovine babesiosis has a re-emerging potential because of the likely loss of herd immunity, while canine babesiosis is rapidly expanding in central and northeastern Europe, its occurrence correlating with the rapid, successful expansion of the ornate dog tick (Dermacentor reticulatus) populations in Europe. Taken together, our analysis of the available reports shows clear evidence of an increasing annual incidence of babesiosis across Europe in both humans and animals that is changing in line with similar increases in the incidence of other tick-borne diseases. This situation is of major concern, and we recommend more extensive and frequent, standardized monitoring using a "One Health" approach.

First Molecular Detection of Bartonella bovis and Bartonella schoenbuchensis in European Bison (Bison bonasus).

Bartonella bacteria infect the erythrocytes and endothelial cells of mammalians. The spread of the Bartonella infection occurs mainly via bloodsucking arthropod vectors. Studies on Bartonella infection in European bison, the largest wild ruminant in Europe, are lacking. They are needed to clarify their role in the maintenance and transmission of Bartonella spp. The aim of this study was to investigate the presence of the Bartonella pathogen in European bison and their ticks in Lithuania. A total of 38 spleen samples from bison and 258 ticks belonging to the Ixodes ricinus and Dermacentor reticulatus species were examined. The bison and tick samples were subjected to ssrA, 16S-23S rRNA ITS, gltA, and rpoB partial gene fragment amplification using various variants of PCR. Bartonella DNA was detected in 7.9% of the tissue samples of European bison. All tick samples were negative for Bartonella spp. The phylogenetic analysis of 16S-23S rRNA ITS, gltA, and rpoB partial gene fragment revealed that European bison were infected by B. bovis (2.6%) and B. schoenbuchensis (5.3%). This is the first report addressing the occurrence of B. bovis and B. schoenbuchensis in European bison in Europe.

Impact of tick-borne Anaplasma phagocytophilum infections in calves of moose (Alces alces) in southern Norway.

The Gram-negative, obligate intracellular tick-transmitted pathogen Anaplasma phagocytophilum can cause acute febrile diseases in humans and domestic animals. The expansion of the tick Ixodes ricinus (Linnaeus, 1758) in northern Europe due to climate change is of serious concern for animal and human health. The aim of the present study was to investigate the impact of A. phagocytophilum infection in moose Alces alces (Linnaeus) calves by evaluating the carcass weights of infected and non-infected animals and examining animal tissues samples for co-infections with either species of Babesia Starcovici, 1893 or bacteria of the genus Bartonella. The carcasses of 68 free-ranging moose calves were weighed by hunters during the hunting seasons from 2014 to 2017 in two regions in southern Norway and spleen samples were collected. Anaplasma phagocytophilum was detected in moose sampled from locations infected with ticks with a prevalence of 82% (n = 46). The carcass weights of A. phagocytophilum-infected calves (n = 46) and non-infected (n = 22) calves were compared. Although the average weight of infected calves (45.6 kg) was lower than that of non-infected calves (46.5 kg), the difference was not statistically significant. Three different variants of the bacterium 16S rRNA gene were identified. The average weight of animals infected with variant I was 49.9 kg, whereas that of animals infected with variant III was 42.0 kg, but the difference was not statistically significant (p = 0.077). Co-infections of A. phagocytophilum with Bartonella spp. or with Babesia spp. were found in 20 and two calves, respectively. A triple infection was found in two calves. Sequence analysis of the 18S rRNA gene of Babesia-positive samples revealed the presence of Babesia cf. odocoilei (Emerson et Wright, 1970). Strains of Bartonella closely related to Bartonella bovis (Bermond, Boulouis, Heller, Laere, Monteil, Chomel, Sander, Dehio et Piemont, 2002) were identified based on phylogenetic analysis of the gltA and rpoB genes. The loss of body mass in moose calves in the tick-infected site was probably influenced by multiple factors.

Bartonella Infections in Cats and Cat Fleas in Lithuania.

Bartonella are vector-borne parasitic bacteria that cause zoonotic infections in humans. One of the most common infections is cat-scratch disease caused by Bartonella henselae and Bartonella clarridgeiae. Cats are the major reservoir for these two species of bacteria, while cat fleas are vectors for the transmission of infection agents among cats. The aim of the present study was to investigate the presence of Bartonella infections in stray and pet cats and in cat fleas in Lithuania. Blood samples were taken from 163 cats presented in pet clinics and animal shelters. A total of 102 fleas representing two species, Ctenocephalides felis and Ctenocephalides canis, were collected from 12 owned cats that live both outdoors and indoors. Bartonella DNA in samples was detected using a nested PCR targeting the 16S-23S rRNA intergenic spacer (ITS) region. Bartonella DNA was detected in 4.9% (8/163) of the cats and 29.4% (30/102) of the fleas. Sequence analysis of the ITS region showed that the cats and fleas were infected with B. henselae, B. clarridgeiae and Bartonella sp., closely related to B. schoenbuchensis. This study is the first report on the prevalence and molecular characterization of Bartonella spp. in cats and cat fleas in Lithuania.

Molecular detection of Babesia spp. in European bison (Bison bonasus) and their ticks.

Babesia spp. are tick-borne haemoparasites that infect a wide range of domestic and wild mammals. Free-ranging ungulates are considered to be important reservoir hosts of Babesia parasites. The European bison (Bison bonasus) is a large and rare ungulate species, reintroduced into the forests of Central Europe after an absence of several decades. Owing to their protected status, studies of tick-borne pathogens in European bison have so far been rare and fragmented. The aim of this study was to investigate the presence of Babesia infection in free-ranging and captive herds of European bison and their ticks. Tissue samples obtained from 37 European bison individuals and 242 ticks belonging to two species, Ixodes ricinus and Dermacentor reticulatus, collected from bison were subjected to PCR analysis of the 18S rRNA gene followed by sequencing. Babesia spp. were detected in 8% of the samples from European bison and in 11% of the ticks. Sequence analysis of partial 18S rRNA gene indicated the presence of B. divergens and B. capreoli in European bison, while B. divergens, B. microti and B. venatorum were detected in ixodid ticks. To the best of authors' knowledge, this is the first molecular detection and characterization of Babesia spp. in European bison and their ticks.

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' strain.

New Records of Bartonella spp. and Rickettsia spp. in Lice Collected from Small Rodents.

Lice are blood-sucking insects that are of medical and veterinary significance as parasites and vectors for various infectious agents. More than half of described blood-sucking lice species are found on rodents. Rodents are important hosts of several Bartonella and Rickettsia species, and some of these bacteria are characterized as human pathogens in Europe. Rodent ectoparasites, such as fleas and ticks, are important vectors of Bartonella spp. and Rickettsia spp., but knowledge about the presence of these bacteria in lice is limited. The aim of this study was to determine the prevalence of Bartonella and Rickettsia bacteria in lice collected from rodents in Slovakia. The ectoparasites were collected from small rodents captured from 2010 to 2015 at four different sites in eastern Slovakia. The presence of Bartonella and Rickettsia species in lice samples was screened by real-time PCR, targeting ssrA and gltA genes, respectively. The molecular characterization of the Bartonella strains was based on sequence analysis of partial rpoB and intergenic spacer (ITS) genes, and of the Rickettsia species on sequence analysis of the gltA gene. A total of 1074 lice of seven species were collected from six rodent species. Bartonella DNA was detected in Hoplopleura affinis (collected from Apodemus agrarius, Apodemus flavicollis, and Myodes glareolus), Polyplax serrata (from A. agrarius), and Hoplopleura sp. (from A. flavicollis). Sequence analysis revealed that the Bartonella strains belonged to the Bartonella coopersplainsensis, Bartonella tribocorum, and Bartonella taylorii genogroups. Rickettsia DNA was detected in H. affinis and P. serrata collected from A. agrarius. Sequence analysis revealed two Rickettsia species: Rickettsia helvetica and Rickettsia sp. The results of the study confirm the presence of Bartonella spp. and Rickettsia spp. in lice collected from rodents.

Prevalence of tick-borne encephalitis virus in questing Dermacentor reticulatus and Ixodes ricinus ticks in Lithuania.

The Baltic States are the region in Europe where tick-borne encephalitis (TBE) is most endemic, with one-third of the European TBE cases detected in Lithuania, Latvia and Estonia. With the Czech Republic and Germany, Lithuania has among the highest population incidences of TBE. Ticks from the Ixodidae family are the main vectors of the TBE virus (TBEV) in Europe. However, there is still a lack of data on the prevalence of TBEV in ticks in different parts of Lithuania. This study analysed the current prevalence of TBEV in the two most common tick species distributed in Lithuania: Ixodes ricinus and Dermacentor reticulatus. Questing I. ricinus (n = 7170) and D. reticulatus (n = 1676) ticks were collected from 81 locations in all ten counties of Lithuania between 2017 and 2019. The presence of TBEV was analysed using a real-time reverse transcription polymerase chain reaction (RT-PCR) and TBEV prevalence in ticks was calculated as the minimum infection rate (MIR). TBEV was detected in the three developmental stages (adults, nymphs and larvae) of I. ricinus and in D. reticulatus adults. The MIR of TBEV in the total sample of I. ricinus was 0.4 % (28/7170) and for D. reticulatus was also estimated to be 0.4 % (6/1676). TBEV-infected ticks were found in 16 locations in seven counties, with MIR ranging from 0.1 % to 1.0 %. The TBEV strains detected belong to the European subtype. To the best of the authors' knowledge, this is the first report on the prevalence of TBEV in unfed D. reticulatus ticks and in I. ricinus unfed larvae in Lithuania.

The prevalence, abundance, and molecular characterization of Lipoptena deer keds from cervids.

The objectives of this study were to investigate the prevalence and abundance of deer keds on various cervids in Lithuania, to molecularly characterize the deer ked species based on mitochondrial COI and 16S rRNA genes, and to compare them with Lipoptena species found in other countries. A total of 11,939 deer keds (Lipoptena cervi and Lipoptena fortisetosa) was collected from the fur of 30 cervids in Lithuania between 2015 and 2019. The values of infestation with deer keds differed among the species of the hosts. Moose and red deer were more frequently infested with L. cervi than with L. fortisetosa, while L. fortisetosa was found more often on roe deer. Phylogenetic analysis of the COI and 16S rRNA genes of five Lipoptena species revealed 110 and 55 variable nucleotide positions, respectively. Among Lithuanian samples, three COI haplotypes of L. cervi and three haplotypes of L. fortisetosa were detected, while there was no variation observed in the 16S rRNA sequences analyzed with one haplotype of L. cervi and one haplotype of L. fortisetosa. This is the first study on L. cervi and L. fortisetosa parasitizing cervids and the first molecular characterization of these deer ked species in Lithuania.

First record of Haemaphysalis concinna (Acari: Ixodidae) in Lithuania.

The relict tick Haemaphysalis concinna Koch, 1844 (Acari: Ixodidae) has high medical and veterinary importance as it is known as a vector or a carrier of various pathogens causing human and animal diseases. Haemaphysalis concinna is a widely distributed species in forests of temperate Eurasia. In Europe, it is found mainly in deciduous or mixed forests near shores of lakes or rivers. However, there is still a lack of knowledge about current distribution, relative density, and other ecological parameters of H. concinna. On June 30th, 2019, one specimen of H. concinna was found on the fur of a hunted roe deer in Lithuania. This is the first record of H. concinna in Lithuania and, to the best of our knowledge, it represents the northernmost distribution point of H. concinna in Europe (22°55'26.5"E/ 55°27'50.7"N).

Associations between different Laelapidae (Mesostigmata: Dermanyssoidea) mites and small rodents from Lithuania.

Associations between species of Laelapidae (Mesostigmata: Dermanyssoidea) mites and small rodents have been studied insufficiently. The aim of this study was to investigate infestation patterns of small rodent species by laelapid mites at six locations in Lithuania. A total of 728 rodents were snap- and live-trapped in various locations during 2013-2016. Eight rodent species were identified, namely Apodemus flavicollis, Apodemus agrarius, Myodes glareolus, Micromys minutus, Mus musculus, Microtus oeconomus, Microtus arvalis and Microtus agrestis. A total of 343 (47.1%) rodents were found to be infested with up to eight species of parasitic mites from the Laelapidae family (n = 1363): Laelaps agilis, Laelaps hilaris, Hyperlaelaps microti, Haemogamasus nidi, Haemogamasus hirsutus, Eulaelaps stabularis, Hirstionyssus sunci and Myonyssus gigas. The dominant species of mite found on rodents was L. agilis (89.1%), found on 43.4% of all hosts. Abundance and mean intensity of infestation with mites varied among species of hosts and were highest for A. flavicollis. We document new geographical and host records for gamasid mites of eight rodent species in Lithuania.

New record of Rickettsia vini in Ixodes lividus ticks from Lithuania.

Ixodes lividus Koch, 1844 (Ixodida: Ixodidae) ticks are parasites specific to the sand martin (Riparia riparia). The distribution range of I. lividus covers Europe and Asia and they are found wherever sand martins nest. Borrelia burgdorferi sensu stricto, Borrelia garinii and Rickettsia vini have recently been detected in I. lividus ticks in Central Europe. The aim of this study was to investigate the presence of Rickettsia spp. (Rickettsiales: Rickettsiaceae) and Borrelia spp. (Spirochaetales: Spirochaetaceae) in I. lividus ticks in Lithuania. A total of 2622 non-engorged I. lividus from two different colonies comprising 40 sand martin nests were collected in Lithuania in 2013 and 2015. Rickettsial DNA was detected in 106 of the 117 pooled larvae samples, in all examined seven nymphs, four females and one male specimen. Sequence analysis of partial gltA, ompA and htrA genes showed 99-100 % identity with the corresponding R. vini sequences deposited in GenBank. None of I. lividus tested was infected with Borrelia burgdorferi sensu lato. This is the first time that the presence of R. vini has been reported in Lithuania.

Prevalence and diversity of Bartonella species in small rodents from coastal and continental areas.

Worldwide, Bartonella infections are known to inflict a wide range of mammals and, within rodents alone, more than 20 Bartonella species have been detected. There is, however, a lack of studies on the presence of Bartonella spp. in rodents in the Baltic region. We analysed 580 individuals belonging to eight small rodent species trapped in coastal and continental areas of Lithuania during 2015-2016. The presence of Bartonella DNA was examined by real-time PCR targeting the ssrA gene. The molecular characterization of the bacteria strains was based on sequence analysis of two housekeeping genes (rpoB, groEL) and the intergenic spacer region (ITS). For the rodents overall, the prevalence of Bartonella spp. was 54.8%, while the prevalence figures for each of the individual species were 8.3% in M. musculus, 15.8% in A. agrarius, 33.3% in M. arvalis, 42.4% in M. glareolus, 53.4% in M. oeconomus, 57.5% in M. minutus, 79.6% in A. flavicollis to 80% in M. agrestis. Sequence analysis revealed that the Bartonella strains belonged to the B. grahamii, B. taylorii, B. rochalimae, B. tribocorum, B. coopersplainsensis and B. doshiae genogroups. The highest Bartonella infection rates and the highest species diversity were both detected in rodents captured in the coastal area. To our knowledge, these are the first reports of the presence of B. coopersplainsensis, B. doshiae and B. tribocorum in Lithuania.