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.

Environment-related and host-related factors affecting the occurrence of lice on rodents in Central Europe.

We studied the effects of environment- (habitat, season) and host-related (sex, body mass) factors on the occurrence of four species of lice (Insecta:Phthiraptera:Anoplura) on six rodent species (Rodentia:Muridae). We asked how these factors influence the occurrence of lice on an individual host and whether different rodent-louse associations demonstrate consistent trends in these effects. We found significant effects of at least one environment-related and at least one host-related factor on the louse occurrence in five of six host-louse associations. The effect of habitat was significant in two associations with the occurrence of lice being more frequent in lowland than in mountain habitats. The effect of season was significant in five associations with a higher occurrence of infestation during the warm season in four associations and the cold season in one association. Host sex affected significantly the infestation by lice in three associations with a higher frequency of infestation in males. Host body mass affected the occurrence of lice in all five associations, being negative in wood mice and positive in voles. In conclusion, lice were influenced not only by the host- but also by environment-related factors. The effects of the latter could be mediated via life history parameters of a host.

Detection of shrew-borne hantavirus in Eurasian pygmy shrew (Sorex minutus) in Central Europe.

Recently, it was found that not only rodents but also shrews are reservoir hosts of hantaviruses. In Central Europe, only Seewis virus, associated with the Eurasian common shrew (Sorex araneus), has been recognized until now. In the present report, tissue samples from shrews belonging to Crocidurinae and Soricinae subfamilies, trapped in Czech Republic, Germany, and Slovakia, were screened for the presence of novel hantaviruses. Three new hantavirus partial L-segment sequences were obtained from pygmy shrews (Sorex minutus) trapped in Czech Republic and Germany. Complete nucleocapsid protein- and glycoprotein precursor-coding S- and M-segment sequences were then determined for the newly recognized hantavirus strains, CZ/Beskydy/412/2010/Sm, CZ/Drahany/420/2010/Sm, and DE/Dürrbach/1912/2009/Sm. Phylogenetic analyses showed that they represent strains of Asikkala virus (ASIV), a novel hantavirus also found in pygmy shrews from Finland. Our study reveals a broad geographic distribution of ASIV across Europe and indicates pygmy shrew as the primary reservoir host. Future studies will have to determine the pathogenic relevance of ASIV.

Broad geographical distribution and high genetic diversity of shrew-borne Seewis hantavirus in Central Europe.

For a long time hantaviruses were believed to be exclusively rodent-borne pathogens. Recent findings of numerous shrew- and mole-borne hantaviruses raise important questions on their phylogenetic origin. The objective of our study was to prove the presence and distribution of shrew-associated Seewis virus (SWSV) in different Sorex species in Central Europe. Therefore, a total of 353 Sorex araneus, 59 S. minutus, 27 S. coronatus, and one S. alpinus were collected in Germany, the Czech Republic, and Slovakia. Screening by hantavirus-specific L-segment RT-PCR revealed specific amplification products in tissues of 49 out of 353 S. araneus and four out of 59 S. minutus. S-segment sequences were obtained for 45 of the L-segment positive S. araneus and all four L-segment positive S. minutus. Phylogenetic investigation of these sequences from Germany, the Czech Republic, and Slovakia demonstrated their similarity to SWSV sequences from Hungary, Finland, Austria, and other sites in Germany. The low intra-cluster sequence variability and the high inter-cluster divergence suggest a long-term SWSV evolution in isolated Sorex populations. In 28 of the 49 SWSV S-segment sequences, an additional putative open reading frame (ORF) on the opposite strand to the nucleocapsid protein-encoding ORF was identified. This is the first comprehensive sequence analysis of SWSV strains from Germany, the Czech Republic, and Slovakia, indicating its broad geographical distribution and high genetic divergence. Future studies have to prove whether both S. araneus and S. minutus represent SWSV reservoir hosts or spillover infections are responsible for the parallel molecular detection of SWSV in both species.

Blood parasites of mound-building mouse, Mus spicilegus Petényi, 1882 (Mammalia, Rodentia).

Mound-building mice, Mus spicilegus, were studied for the blood parasites in Eastern Slovakia, vicinity Kechnec village near Kosice town (Kosická kotlina basin, 21 degrees 14' E, 48 degrees 33' N) during years 2002-2005. Overall, 251 specimens were examined. The parasites were detected using microhematokrit centrifugation technique and on the Giemsa's method stained blood smears and light microscopy. The parasites were found in 3.57% of specimens; 1.20% of mice were infected with Bartonella sp., 2.39% were infected with Babesia piroplasms. No Hepatozoon hemogregarines and trypanosomes were observed. The intensity of infection with Bartonella was low, less than 0.01% of erythrocytes were invaded, the percent of the erythrocytes with Babesia sp. was less than 0.01%. The morphological description and measurements of parasites were made using the "Analysis" software combined with a video camera and a microscope. The mean size of Bartonella sp. bacteria's were 0.8 x 0.3 microm, range 0.4-1.5 x 0.1-0.9 microm, Babesia sp. occurred in pear-shaped and ring-like forms, 1.00-1.27 microm in diameter, and 0.98-1.27 microm in size, respectively. The regular form of four cells--"maltese cross" was not noticed. This is the first record infection of Mus spicilegus with blood parasites.

Small mammals (Insectivora, Rodentia) as a potential source of chlamydial infection in East Slovakia.

The presence of antibodies against Chlamydophila (Chlamydia) psittaci in small mammals (Insectivora, Rodentia) in the region of East Slovakia are presented. The hosts were caught in several areas of Slovakia in habitats with different levels of anthropogenic disturbance. Research was carried out during 2000-2002. The authors examined 1,947 sera coming from 4 insectivore and 10 rodent species. Each serum was examined by micromethod of complement binding reactions using antigen Chlamydophila (Chlamydia) psittaci. Chlamydial infections were found in 251 individuals (prevalence 12.9 %) of 8 mammal species. The antichlamydial antibodies were proved at levels ranging from 1:32-1:1024. The highest prevalence of antibodies was detected in the most abundant rodent species Apodemus microps (14.8 %), Apodemus agrarius (13.9 %), Apodemus flavicolis (12.4 %), Microtus arvalis (12 %), and Clethrionomys glareolus (10.9 %). Positive hosts were registered in all studied localities. Testing of prevalence values in the individual research years confirmed significant changes. Our results showed that small mammals probably play an important role in the circulation of chlamydiae in nature.

Anti-Borrelia antibodies in rodents: important hosts in ecology of Lyme disease.

Data presented in this study focuses on the presence of anti-Borrelia antibodies in small mammals from Eastern Slovakia during 2000-2003. The total seropositivity observed was 18.78% in rodents. Amongst all species, the total seroprevalence in Apodemus flavicolis was the highest (20.87%), followed by Apodemus agrarius (19.58%) and Clethrionomys glareolus (11.11%). However, the prevalence in Apodemus flavicolis during the year 2000-2001 was higher (26.72%), which reduced to 10.60% in 2002-2003. To compare the year range of seroprevalence in other small mammals was not feasible due to the small sample number. Area-wise distribution of anti-Borrelia antibodies was even (18.75% to 20%) in this study, except in the Bot'any province (0%). This confirms the equal distribution of Borrelia spirochetes in the other 3 localities. Prevalence of anti-Borrelia antibodies during summer was significantly higher than during autumn and early spring. The overall study also reviews the importance of small mammals in Lyme disease ecology.