European Rodents Research Articles

Marsupials (Herpetotheriids) from the late Palaeogene of south-east Serbia

Three marsupial species are present in Palaeogene assemblages from south-eastern Serbia, Amphiperatherium minutum (Aymard, 1846), A. exile (Gervais, 1848–1852) and a species of Peratherium Aymard, 1850. These species are common in the late Eocene and early Oligocene of Western and Central Europe and their presence in South-Eastern Europe indicate that by the end of the Eocene, the eastwards dispersal of these marsupial species was already a fact. The presence of the same marsupial species in Serbia and in Western Europe is surprising, as the difference in composition between the Serbian and Western European rodent faunas indicates a limited faunal exchange between these areas in the late Eocene and early Oligocene. In marsupials, the stylar shelf of the upper molars shows a large morphological variation. A lesser-known variation is the presence of a transverse crista in the stylar shelf. These occur in various marsupial species, but so far were not known to be present in Amphiperatherium minutum. In our opinion, the large morphological variation of the stylar shelf is thus better not used to define a new species.

A buprenorphine depot formulation provides effective sustained post-surgical analgesia for 72 h in mouse femoral fracture models

Adequate pain management is essential for ethical and scientific reasons in animal experiments and should completely cover the period of expected pain without the need for frequent re-application. However, current depot formulations of Buprenorphine are only available in the USA and have limited duration of action. Recently, a new microparticulate Buprenorphine formulation (BUP-Depot) for sustained release has been developed as a potential future alternative to standard formulations available in Europe. Pharmacokinetics indicate a possible effectiveness for about 72 h. Here, we investigated whether the administration of the BUP-Depot ensures continuous and sufficient analgesia in two mouse fracture models (femoral osteotomy) and could, therefore, serve as a potent alternative to the application of Tramadol via the drinking water. Both protocols were examined for analgesic effectiveness, side effects on experimental readout, and effects on fracture healing outcomes in male and female C57BL/6N mice. The BUP-Depot provided effective analgesia for 72 h, comparable to the effectiveness of Tramadol in the drinking water. Fracture healing outcome was not different between analgesic regimes. The availability of a Buprenorphine depot formulation for rodents in Europe would be a beneficial addition for extended pain relief in mice, thereby increasing animal welfare.

Seasonal dynamics of the wild rodent faecal virome.

Viral discovery studies in wild animals often rely on cross-sectional surveys at a single time point. As a result, our understanding of the temporal stability of wild animal viromes remains poorly resolved. While studies of single host-virus systems indicate that host and environmental factors influence seasonal virus transmission dynamics, comparable insights for whole viral communities in multiple hosts are lacking. Utilizing noninvasive faecal samples from a long-term wild rodent study, we characterized viral communities of three common European rodent species (Apodemus sylvaticus, A. flavicollis and Myodes glareolus) living in temperate woodland over a single year. Our findings indicate that a substantial fraction of the rodent virome is seasonally transient and associated with vertebrate or bacteria hosts. Further analyses of one of the most common virus families, Picornaviridae, show pronounced temporal changes in viral richness and evenness, which were associated with concurrent and up to ~3-month lags in host density, ambient temperature, rainfall and humidity, suggesting complex feedbacks from the host and environmental factors on virus transmission and shedding in seasonal habitats. Overall, this study emphasizes the importance of understanding the seasonal dynamics of wild animal viromes in order to better predict and mitigate zoonotic risks.

Living in a changing world: Physiological and behavioural flexibility of juvenile Garden Dormice

Heterothermy, or torpor, allows individuals to save energy by active reduction of metabolism and decreased body temperature. Social thermoregulation or huddling allows individuals to minimize energy needs while maintaining a relatively high body temperature. The Garden Dormouse (Eliomys quercinus), a highly endangered European rodent, uses both strategies of torpor and huddling, which are particularly beneficial for young individuals to cope with environmental fluctuations. Since juveniles are an important component of population renewal, determining the flexibility of energy-saving strategies and its consequences on somatic integrity of young individuals is key to protecting this species. We measured individual torpor patterns, body mass gain and structural growth, and assessed telomere dynamics in male and female juvenile Garden Dormice according to housing condition (singly or groups of four individuals) and food availability (fed ad-libitum or energy-limited) before and during hibernation. During development, juveniles used more frequent, longer and deeper torpor when housed singly than in groups. Torpor was encouraged by lower food availability. Juveniles showed similar body mass gain and growth, irrespective of experimental conditions. During hibernation, huddling dormice showed similar hibernating patterns and mass loss compared to single individuals regardless of food availability. Telomere lengthened across all individuals before hibernation, with larger elongation in males, which also experienced greater telomere loss than females during hibernation. We conclude that the use of torpor and huddling allows juvenile Garden Dormice to cope successfully with energetic bottlenecks, without major effects on somatic integrity. Future studies will need to confirm such findings under natural environmental conditions.

Combining citizen science and conservation genomics to reveal the causes of rapid population decline in the Garden Dormouse (Eliomys quercinus)

The Garden Dormouse (Eliomys quercinus) is a native European rodent species that has suffered extensive range contraction and severe population decline across large parts of its distribution in Central and Eastern Europe and is now considered extinct in several countries. At the same time, contrasting population dynamics are observed in its western distribution, where Garden Dormice are regionally abundant and even occupy habitats in the built environment. Reasons for these opposing range dynamics and drastic declines remain unknown. We therefore started a long-term cooperation project, joining the efforts of different research institutions, conservation NGOs and citizen scientists to investigate the underlying causes. Here, we present the results of a RADseq analysis of 103 samples collected from several parts of the European species range to infer regional phylogeography and delineate different conservation units. Preliminary genomic data show the existence of highly divergent genetic lineages, even on a local scale. Based on these findings, we have developed a reduced SNP panel, which allows for lineage discrimination, assessment of population structure and identification of individuals from non-invasively collected samples. The panel is now routinely applied in Garden Dormouse conservation, e.g., when selecting appropriate locations to release displaced Garden Dormice found accidentally or to identify isolated Garden Dormouse populations in need of improved habitat connectivity.

The beginning of the adaptive radiation of Theridomorpha (Rodentia) in Western Europe: morphological and phylogenetic analyses of early and middle Eocene taxa; implications for systematics

This paper provides a revision of the early and middle Eocene European rodents previously referred to as Ischyromyoidea, including taxa considered to be at the origin of the Theridomorpha. The use of an accurate dental terminology and a better understanding of the size and shape of their infra-orbital foramen (i.o.f.) led us to a substantial revision of this group, which allowed to better characterize them and to appreciate their variability. On these bases, phylogenetic analyses (cladistic and standard Bayesian approaches) of early Ypresian to late Priabonian European rodent species were undertaken in order to highlight the root of the early Theridomorpha and its content. The phylogeny was established based on 343 characters (338 dental) through 45 early Paleogene taxa. The ingroup included on one hand a few North American genera (Reithroparamys, Microparamys, and Acritoparamys) and European ones (Eogliravus, Ailuravus, Corbarimys, Meldimys, Euromys, Plesiarctomys, and Pseudoparamys) considered until now as being related with the North American superfamily Ischyromyoidea. On the other hand, it included genera close to the root of the Theridomorpha (Sparnacomys, Pantrogna, and Hartenbergeromys) and early Theridomyoidea (Masillamys, Protadelomys, and some Pseudosciuridae). The phylogenetic results obtained via the two distinct reconstruction approaches are consistent in virtually all relationships. The proposed systematics here derives from these phylogenetic results. This phylogenetic context led us to change the suprafamilial, familial, subfamilial or generic attribution of several species. Characters of Theridomorpha, like the obliquely developed postprotocristid allied with the occurrence of a metalophulid I, have been found in genera previously considered as Ischyromyidae (Pseudoparamys, Euromys, Sparnacomys, Meldimys, Pantrogna, and Hartenbergeromys) as well as the large i.o.f., when preserved (Pseudoparamys, Hartenbergeromys, and Masillamys). Based on these morphological observations and new phylogenetic considerations, the content of the Theridomorpha clade is here enlarged, thereby extending back the first theridomorph radiations to the early Eocene. Aside, a new taxon (Reinomys rhomboides gen. and sp. nov.) is described from Avenay. In addition, a new genus, Auroremys, is created for the species subita (Comte et al., 2012) from Chery-Chartreuve.

Hepatozoon in Eurasian red squirrels Sciurus vulgaris, its taxonomic identity, and phylogenetic placement.

Adeleorid apicomplexan parasites of the genus Hepatozoon Miller, 1908 are broadly distributed among the rodents. Broader molecular data on Hepatozoon from Palaearctic squirrels are necessary for evaluation of diversity and origin of Hepatozoon in Eurasian red squirrel Sciurus vulgaris populations, considering ongoing invasion by Gray squirrel S. carolinensis. Our report brings a set of molecular data from a population of S. vulgaris in the Czech Republic, non-invaded by any invasive squirrel species. Cadavers of 41 Eurasian red squirrels were examined using nested PCR targeting 18S rRNA gene; 30 animals tested positive for the presence of Hepatozoon spp. DNA in at least one tissue. Phylogenetic analysis of obtained sequence types revealed relatedness to sequences of Hepatozoon sp. from S. vulgaris from Spain and the Netherlands, forming a sister clade to Hepatozoon isolates from other European rodents. The fact that all available 18S rRNA gene sequences form a monophyletic clade is interpreted as a presence of a single Hepatozoon species in S. vulgaris in continental Europe, most probably Hepatozoon sciuri. The presented molecular data on the Hepatozoon from European squirrels provides a basis for future studies on possible exchange of Hepatozoon species between Eurasian red and gray squirrels.

Patterns of activity rhythms of invasive coypus Myocastor coypus inferred through camera-trapping.

Studies on activity rhythms are pivotal for the management of invasive alien species, as they provide basic insights into species basic ecology and may increase the success of control programs. The coypu Myocastor coypus, introduced from South America for fur farms, has become one of the most invasive rodents in Europe. Introduced coypus may affect crop productions, as well as natural vegetation and the breeding success of wading birds. In this study, we examined activity data collected through intensive camera-trapping in three Italian areas, including two natural areas in Northern and Central Italy, and a suburban area in Central Italy. Coypus were mostly diurnal in areas characterised by low predator pressure and, at night, they are mostly active in bright moonlight. Conversely, where predators, human pressure or numerical control programmes are present, coypus remarkably shift their behaviour towards crepuscular and night hours. In these last areas, nocturnal activity increased as moonlight decreased, possibly to reduce predation risk or encounters with humans. Where winter temperature are low, diurnal habits may have developed as a physiological adaptation and a strategy to preserve energy, potentially achieving a cost/effective thermal balance.Electronic supplementary materialThe online version of this article (10.1007/s42991-020-00052-8) contains supplementary material, which is available to authorized users.

Multi-Locus Genetic Identification of a Newly Discovered Population Reveals a Deep Genetic Divergence in European Blind Mole Rats (Rodentia: Spalacidae: Nannospalax)

A new population of blind mole rat (genus Nannospalax) was discovered near the town of Albertirsa in north-central Hungary. We used newly designed primers to specifically amplify the whole mitochondrial cytochrome-b region and two nuclear DNA regions. Based on the most comprehensive taxonomic sampling to date, we compared this population with several other European blind mole-rat taxa. The results from both mitochondrial and nuclear regions have unequivocally placed the Albertirsa population into the monophyletic group of the Vojvodina blind mole rat (N. (leucodon) montanosyrmiensis), which turned out to be a sister clade to all other molecularly studied European Nannospalax. This study not only identified the fourth known population of an extremely rare rodent taxon but also calls for a taxonomic revision of European lesser blind mole rats (N. leucodon superspecies) to systematically evaluate the genetic structure of their populations and to understand the complex evolutionary history of these European rodents. The occurrence of the Vojvodina blind mole rat at this northern location helps to clarify the distribution area of this heavily data-deficient taxon. As currently understood, this lineage predominantly occurs in sandy grasslands of the Danube–Tisza Interfluve in Hungary and Serbia. Its distribution range and phylogenetic structure might reflect the importance of potential biogeographical barriers (e.g. large rivers) that shaped the blind mole rats' allopatric or peripatric speciation.

Gimme shelter: The effect of rocks and moonlight on occupancy and activity pattern of an endangered rodent, the garden dormouse Eliomys quercinus

Information on spatial behaviour and temporal activity patterns is paramount for the conservation of animal species. This is particularly true for endangered taxa that are threatened by ongoing climatic and environmental changes. The garden dormouse Eliomys quercinus is a native European rodent (family Gliridae), whose populations are declining throughout the Continent. Notwithstanding this, neither International nor National laws explicitly require mandatory monitoring of populations. As a result, compelling information on the spatiotemporal behaviour of dormouse is lacking. We aimed to fill this gap by investigating occupancy patterns in relation to environmental features and activity rhythms in relation to moonlit nights in an Alpine population of dormouse within the Stelvio National Park, northern Italy. Data were collected between May and October 2015. Twenty camera-traps were deployed in a 500 ha coniferous forest, using a random tessellation approach; camera trap data were analyzed with occupancy models and kernel smoothers. Camera-traps provided a reliable assessment of the presence of the garden dormouse, with only 1 % of false absence and a high detection probability (68 %). The occurrence of the garden dormouse was positively influenced by the percentage of rock coverage on the ground. The species showed a strictly nocturnal behaviour, with an activity peak before midnight, negatively related to moonlit nights. The use of rocky areas for nesting, shelter site and thigmotactic movements and moonlight avoidance may represent adaptations of the garden dormouse to avoid predation risk. Our results shed some light on the habitat requirement of a poorly known, near-threatened species, and provide baseline information for future monitoring and conservation activities.

Distribution of Neoehrlichia mikurensis in Ixodes ricinus ticks along the coast of Norway: The western seaboard is a low‐prevalence region

Neoehrlichia mikurensis is a tick-borne pathogen widespread among ticks and rodents in Europe and Asia. A previous study on Ixodes ricinus ticks in Norway suggested that N.mikurensis was scarce or absent on the south-west coast of Norway, but abundant elsewhere. The aim of this study was to further investigate the prevalence and distribution of N.mikurensis along the western seaboard of Norway in comparison with more eastern and northern areas. The second aim of the study was to examine seasonal variation of the bacterium in one specific location in the south-eastern part of Norway. Questing I.ricinus were collected from 13 locations along the coast of Norway, from Brønnøysund in Nordland County to Spjaerøy in Østfold County. In total, 11,113 nymphs in 1,113 pools and 718 individual adult ticks were analysed for N.mikurensis by real-time PCR. The mean prevalence of N.mikurensis in adult ticks was 7.9% while the estimated pooled prevalence in nymphs was 3.5%. The prevalence ranged from 0% to 25.5%, with the highest prevalence in the southernmost and the northernmost locations. The pathogen was absent, or present only at low prevalence (<5%), at eight locations, all located in the west, from 58.9°N to 64.9°N. The prevalence of N.mikurensis was significantly different between counties (p<.0001). No significant seasonal variation of N.mikurensis prevalence was observed in the period May to October 2015. Our results confirm earlier findings of a low prevalence of N.mikurensis in the western seaboard of Norway.

Reintroducing beavers Castor fiber to Britain: a disease risk analysis

Abstract Eurasian beavers Castor fiber are potential hosts for a range of infectious diseases and parasites, including those typical of common European rodents. A number of infectious organisms are potentially zoonotic and may be notifiable under animal health legislation. The official trial beaver reintroductions to Scotland, the retrospectively licensed releases in England, and the increasingly obvious presence of large numbers of unlicensed illegally released animals have highlighted potential disease risks. We aimed to conduct a disease risk analysis, based on peer reviewed publications, for selection and health screening of Eurasian beavers prior to release into the wild in Britain. Adapted from the International Union for the Conservation of Nature’s ‘Guidelines for Disease Risk Analysis’, a four‐step process was used to formulate a disease risk analysis: 1) problem description; 2) hazard identification based on literature review; 3) risk assessment, which resulted in categorisation of pathogens into low, medium, and high risk; and 4) risk management: identification of mitigating measures, followed by risk re‐evaluation in light of the reported effectiveness of the mitigation measures. The highest‐risk pathogens identified in the literature review process included: parasites, specifically Cryptosporidium parvum, Echinococcus multilocularis, Eimeria spp., Fasciola hepatica, Giardia spp., Trichinella britovi; bacteria, specifically Escherichia coli, Franciscella tularensis, Mycobacterium avium, Salmonella spp., Yersinia spp.; a fungus Chrysosporium parvum (Emmonsia parva); and terrestrial rabies virus. Most could be mitigated by sourcing beavers from Britain. The rest could be mitigated by pre‐release testing procedures that are already established. The risk of introducing significant disease to humans, domestic animals, or wildlife by releasing into the wild in Britain a beaver that was captive‐bred in Britain or a wild beaver from Scotland, based on the current evidence of disease incidence, and assuming the use of robust, peer reviewed, pre‐release health screening techniques, can be viewed as low.

Discovery of novel highly divergent RNA viruses in European rodents and rabbits

Although RNA viruses are likely to exist in every species of cellular life, our knowledge of their biodiversity and evolution has been focused on those that can cause disease in humans and in economically important animals and plants. Recently published studies, however, have shown that every healthy organism can carry viruses. In this study, we present the unbiased discovery of highly divergent RNA viruses in European rodents and rabbits using Next Generation Sequencing (NGS). Tissue samples from different organs were collected from UK rodents and French rabbits and were initially screened for viruses with PCR. Following up on preliminary data, the positive samples were sent for NGS to acquire full genomes and perform unbiased virus discovery. Our findings encompass a number of novel viruses including astrovirus, rotavirus A, hantavirus, picornavirus, coronavirus, paramyxovirus etc. Among the novel viruses, the picornavirus interestingly showed 50 % and 34 % similarity to the closest relative for the non-structural and the structural protein respectively at amino acid level, suggesting a potentially novel genus within the family. This study shows the presence of highly divergent RNA viruses in European rodents and rabbits enriching thus our current knowledge of their origin and evolution. Finally, this study shows that these animal species can be the reservoirs of RNA viruses notorious for cross-species transmission, increasing the risk of a spill-over to humans or livestock.

Shared Common Ancestry of Rodent Alphacoronaviruses Sampled Globally.

The recent discovery of novel alphacoronaviruses (alpha-CoVs) in European and Asian rodents revealed that rodent coronaviruses (CoVs) sampled worldwide formed a discrete phylogenetic group within this genus. To determine the evolutionary history of rodent CoVs in more detail, particularly the relative frequencies of virus-host co-divergence and cross-species transmission, we recovered longer fragments of CoV genomes from previously discovered European rodent alpha-CoVs using a combination of PCR and high-throughput sequencing. Accordingly, the full genome sequence was retrieved from the UK rat coronavirus, along with partial genome sequences from the UK field vole and Poland-resident bank vole CoVs, and a short conserved ORF1b fragment from the French rabbit CoV. Genome and phylogenetic analysis showed that despite their diverse geographic origins, all rodent alpha-CoVs formed a single monophyletic group and shared similar features, such as the same gene constellations, a recombinant beta-CoV spike gene, and similar core transcriptional regulatory sequences (TRS). These data suggest that all rodent alpha CoVs sampled so far originate from a single common ancestor, and that there has likely been a long-term association between alpha CoVs and rodents. Despite this likely antiquity, the phylogenetic pattern of the alpha-CoVs was also suggestive of relatively frequent host-jumping among the different rodent species.

Distribution and space use of seed-dispersing rodents in central Pyrenees: implications for genetic diversity, conservation and plant recruitment.

The function and conservation of many forest ecosystems depend on the distribution and diversity of the community of rodents that consume and disperse seeds. The habitat preferences and interactions are especially relevant in alpine systems where such granivorous rodents reach the southernmost limit of their distribution and are especially sensitive to global warming. We analyzed the community of granivorous rodents in the Pyrenees, one of the southernmost mountain ranges of Europe. Rodent species were identified by DNA with particular attention to the Apodemus species, which are prominent seed-dispersing rodents in Europe. We confirmed for the first time the presence of the yellow-necked mouse, Apodemus flavicollis, in central Pyrenees, a typical Eurosiberian species that reaches its southernmost distribution limit in this area. We also found the wood mouse, Apodemus sylvaticus, a related species more tolerant to Mediterranean environments. Both rodents were spatially segregated by altitude. A. sylvaticus was rare at high altitudes, which might cause the genetic differentiation between populations of the different valleys reported here. We also found other seed consumers like dormice, Elyomis quercinus, and voles, Myodes glareolus, with marked habitat preferences. We suggest that population isolation among valleys may increase the genetic diversity of rodents, like A. sylvaticus. We also highlight the potential threat that global warming may represent for species linked to high-altitude refuges at the southern edge of its distribution, like Apodemus flavicollis. Finally, we discuss how this threat may have a dimension in the conservation of alpine forests dispersed by these rodent populations.

First Record of Gyrabascus (Digenea, Pleurogenidae) from Dromiciops bozinovici D'Elia et al., 2016 (Marsupialia: Microbiotheriidae) in Chile and its Phylogenetic Relationships

Members of the digenean genus Gyrabascus have been reported from bats and rodents in Europe, Southeast Asia, North America, and most recently from the marsupial, Monito del Monte (Dromiciops gliroides), in the Argentinian part of Patagonia. We have found digeneans belonging to Gyrabascus in a different species of Monito del Monte (Dromiciops bozinovici) from Chile. This is the first record of Gyrabascus from D. bozinovici and only the second report of Gyrabascus in a marsupial. Although these specimens are morphologically very similar to Gyrabascus dromiciops, we refrained from identifying our material as G. dromiciops due to insufficient specimen quality and the presence of some morphological differences, leaving open the possibility that the specimens from Chile may potentially represent a different, new species. Both morphological characteristics and molecular phylogenetic analysis using partial sequence of nuclear large ribosomal subunit gene (28S) confirmed the systematic position of these specimens and allowed us to discuss their phylogenetic relationships.

Distribution and status of the declining garden dormouseEliomys quercinus

AbstractThe garden dormouseEliomys quercinus, a native European rodent species, suffered a significant contraction in its geographical range in the last few decades. The species has disappeared from large parts of central and eastern Europe and is considered extinct in some countries.I reviewed the information available on the occurrence and distribution of the species in 26 countries where it was previously reported. Present and past introductions outside its native range were also summarised.The garden dormouse is considered extinct in Lithuania, Finland, and Slovakia, probably extinct in Belarus, and present with single populations in the Netherlands, Poland, and Slovenia; in Slovakia, however, monitoring is necessary to verify recent records. The species is rare and localised in Austria, Ukraine, Romania, and Croatia and is in regression in Germany, Flanders (Belgium), Czech Republic, Latvia, and Estonia. In 2015, the garden dormouse occupied 49% of its 1978 geographical range and 67% of its 2008 range.South‐western Europe is the stronghold of the species; it is still common in large parts of Portugal, Spain, France, and Italy. However, there are indications that also in these countries, the species is locally declining.Present knowledge cannot explain the extensive regression of the garden dormouse's range in central and eastern Europe. Probably, it is the result of the interaction of different factors, acting locally and at a large scale, and related to specific ecological requirements of the species.There is a strong need for research to determine the reasons for the dramatic population and geographical range contraction of the garden dormouse. Meanwhile, it is important to monitor this species and to identify appropriate conservation measures.

Did the Quaternary climatic fluctuations really influence the tempo and mode of diversification in European rodents?

The objective of this study was to establish whether the Quaternary climatic fluctuations influenced the tempo and mode of diversification in European rodents. Our case study is the subgenus Microtus (Terricola) distributed from western Europe to the Caucasus. Mitochondrial cytochrome b gene sequences from several representatives of all the species were used to generate maximum-likelihood and Bayesian phylogenetic trees, to estimate divergence times, to identify biogeographic ancestral areas and to study the rate of diversification. Results showed that phylogenetic tree topologies were similar to previous published studies but with a better resolution at some nodes. The origin of Microtus (Terricola) is dated back to approximately 4.05 Myr in the Early Pliocene, and molecular dating for most Terricola species corresponds to several glacial periods of the Pleistocene. Results of the biogeographic ancestral area reconstruction suggest that Microtus (Terricola) diversified from the Caucasus/Turkey/Iran area through western Europe. Several periods of diversity variation were highlighted as follows: two period of diversity increase, between 3 and 2 Myr, and after 1 Myr; two periods of diversity decrease, before 3 Myr, and between 2 and 1 Myr. The diversification rate of Microtus (Terricola) was 0.353 ± 0.004 event/Myr, a rate similar to that of the Muridae family. To conclude, although the Pleistocene glacial conditions had an impact on the speciation events, the Quaternary does not appear however as a period with an exceptional rate of diversification for European rodents.

Molecular detection and phylogenetic analysis of Hepatozoon spp. in questing Ixodes ricinus ticks and rodents from Slovakia and Czech Republic

By amplification and sequencing of 18S rRNA gene fragments, Hepatozoon spp. DNA was detected in 0.08% (4/5057) and 0.04% (1/2473) of questing Ixodes ricinus ticks from Slovakia and Czech Republic, respectively. Hepatozoon spp. DNA was also detected in spleen and/or lungs of 4.45% (27/606) of rodents from Slovakia. Prevalence of infection was significantly higher in Myodes glareolus (11.45%) than in Apodemus spp. (0.28%) (P < 0.001). Sequencing of 18S rRNA Hepatozoon spp. gene amplicons from I. ricinus showed 100% identity with Hepatozoon canis isolates from red foxes or dogs in Europe. Phylogenetic analysis showed that at least two H. canis 18S rRNA genotypes exist in Slovakia of which one was identified also in the Czech Republic. The finding of H. canis in questing I. ricinus suggests the geographical spread of the parasite and a potential role of other ticks as its vectors in areas where Rhipicephalus sanguineus is not endemic. Sequencing of 18S rRNA gene amplicons from M. glareolus revealed the presence of two closely related genetic variants, Hepatozoon sp. SK1 and Hepatozoon sp. SK2, showing 99-100% identity with isolates from M. glareolus from other European countries. Phylogenetic analysis demonstrates that 18S rRNA variants SK1 and SK2 correspond to previously described genotypes UR1 and UR2 of H. erhardovae, respectively. The isolate from Apodemus flavicollis (Hepatozoon sp. SK3b) was 99% identical with isolates from reptiles in Africa and Asia. Further studies are necessary to identify the taxonomic status of Hepatozoon spp. parasitizing rodents in Europe and the host-parasite interactions in natural foci.

Discovery of Novel Alphacoronaviruses in European Rodents and Shrews.

Eight hundred and thirteen European rodents and shrews encompassing seven different species were screened for alphacoronaviruses using PCR detection. Novel alphacoronaviruses were detected in the species Rattus norvegicus, Microtus agrestis, Sorex araneus and Myodes glareolus. These, together with the recently described Lucheng virus found in China, form a distinct rodent/shrew-specific clade within the coronavirus phylogeny. Across a highly conserved region of the viral polymerase gene, the new members of this clade were up to 22% dissimilar at the nucleotide level to the previously described Lucheng virus. As such they might represent distinct species of alphacoronaviruses. These data greatly extend our knowledge of wildlife reservoirs of alphacoronaviruses.