Tick-borne encephalitis virus (TBEV) is the most important causative agent of arboviral infection in Europe, causing neurologic symptoms. The incidence of the disease has greatly increased over the past decades, and in the meantime, some changes in spatial distribution of TBE cases have been observed. Therefore, it is important to recognize the distribution of endemic areas, to use preventive measures successfully. In this study, rodents from all over Slovenia were evaluated as suitable sentinels for TBEV distribution. Rodents from four species (Myodes glareolus, Apodemus flavicollis, Apodemus sylvaticus, and Apodemus agrarius) were screened for the presence of TBEV antibodies with immunofluorescence assay; the antibodies were detected in 5.9% of sera. The prevalence of infection varied according to the rodent species and according to the region of trapping. Select rodents were also screened for the presence of TBEV RNA in several organs. Both analyses showed higher rate of infection in bank voles, which also produced higher titers of anti-TBEV antibodies and a higher TBEV RNA viral load compared with mice. The regional prevalence of infection in rodents can be correlated with the incidence of disease. Molecular results indicate that the virus can be detected in the organs of the rodents for longer periods, indicating prolonged infections of the rodent hosts by the virus. Rodents can therefore be used as a useful indicator of the circulation of TBEV in an area.

Genetic analysis was performed of wild-type (wt) Dobrava hantavirus (DOB) strains from Slovenia, the country where the virus was first discovered and where it was found to cause haemorrhagic fever with renal syndrome (HFRS), with a fatality rate of 12 %. Two hundred and sixty mice of the genus Apodemus, trapped in five natural foci of DOB-associated HFRS during 1990-1996, were screened for the presence of anti-hantavirus antibodies and 49 Apodemus flavicollis and four Apodemus agrarius were found to be positive. RT-PCR was used to recover partial sequences of the wt-DOB medium (M) and small (S) genome segments from nine A. flavicollis and one A. agrarius. Sequence comparison and phylogenetic analysis of the Slovenian wt-DOB strains revealed close relatedness of all A. flavicollis-derived virus sequences (nucleotide diversity up to 6 % for the M segment and 5 % for the S segment) and the geographical clustering of genetic variants. In contrast, the strain harboured by A. agrarius showed a high level of genetic diversity from other Slovenian DOB strains (14 %) and clustered together on phylogenetic trees with other DOB strains harboured by A. agrarius from Russia, Estonia and Slovakia. These findings suggest that the DOB variants carried by the two species of Apodemus in Europe represent two distinct genetic lineages.

Recent bioacoustic investigations have shown that Cicadetta montana Scopoli 1772 is a complex of morphologically similar sister species that are best characterized by their song patterns. At the type locality of C. montana, only mountain cicadas with simple, long lasting song phrases were heard, recorded and collected. Therefore, we have good reasons to suggest that this type of song is characteristic for C. montana s. str. Boulard described a song of C. montana from France with phrases composed of a long and a short echeme; this type of song is characteristic for cicadas morphologically corresponding to C. montana var. brevipennis Fieber 1876; we suggest to raise this taxon to species level. On the basis of specific song, Puissant and Boulard described C. cerdaniensis from Pyrénées. A similar case was the discovery and description of C. montana macedonica Schedl 1999 from Macedonia; since these Macedonian cicadas are sympatric with at least two other cryptic species in the C. montana group and molecular investigations showed substantial genetic differences between C. macedonica and C. montana or C. brevipennis, we conclude that this taxon should also be raised to species level. Songs of closely related C. podolica and Korean mountain cicada are presented as well.

Recent acoustic studies have revealed that Cicadetta montana (Scopoli, 1772), which was once thought to be a single widespread Palaearctic cicada species, is actually a complex of many taxa. Although some song patterns are very distinct, others comprise groups of closely related species, as in the case of Cicadetta cerdaniensis Puissant & Boulard, 2000, Cicadetta cantilatrix Sueur & Puissant, 2007, and Cicadetta anapaistica Hertach, 2011. Seven spatially or behaviourally isolated metapopulations belonging to this song group from Italy and Switzerland were detected and investigated using acoustic, molecular, and morphological methods. Taxonomic decisions in this group are challenging because of a lack of truly diagnostic morphological characters, variously coloured morphs, qualitatively intermediate song patterns in contact zones, and strong temperature dependence of song‐duration characters. Molecular genetic studies suggest rapid speciation resulting in incomplete lineage sorting and introgression. It is only by using multiple sources of data that species can be delimited. The new species Cicadetta sibillae sp. nov. and the new subspecies Cicadetta anapaistica lucana ssp. nov. were described using the microstructure of the male calling songs. Cicadetta sibillae sp. nov. occurs from southern Switzerland to central Italy, and is the most abundant cicada in the Northern Apennine. Cicadetta anapaistica lucana ssp. nov. is endemic to a small southern Italian distribution range, and seems to be threatened by habitat loss and fragmentation. There is strong evidence that current distribution patterns and phylogenetic relationships of the Cicadetta cerdaniensis group are linked to speciation events in Pleistocene glacial refugia in the Italian, Iberian, and Balkan peninsulas. © 2015 The Linnean Society of London

A novel spotted fever group Rickettsia was found in Haemaphysalis sulcata ticks collected from sheep and goats in Croatia in 2006. At the same time, a genetically identical organism was coisolated with the embryonic cell line CCE3 obtained from the soft tick Carios capensis in Georgia, USA. In this study, further phenotypic and genotypic characteristics of the novel rickettsial strain present in H. sulcata ticks were investigated. Based on the cultivation of bacteria in mosquito and Vero cell cultures, the presence of rickettsiae in tick tissues and cell cultures [confirmed by transmission electron microscopy (TEM)] and the amplification and sequencing of five rickettsial genes, it was demonstrated that the novel Rickettsia strain fulfils the criteria to be classified as a novel species. The name Rickettsia hoogstraalii sp. nov. is proposed for the new strain. Rickettsia hoogstraalii sp. nov., an obligately intracellular bacterium, was grown in Vero cells and arthropod CCE3, ISE6 and C6/36 cell lines. The morphology of the cells of the novel species was typical of SFG rickettsiae. The small coccobacillary appearance of the bacteria was apparent with light microscopy. A Gram-negative bacterial cell wall and a cytoplasmic membrane separated by a narrow periplasmic space were visible by TEM. To date, Rickettsia hoogstraalii sp. nov. has been isolated from two species of ticks, H. sulcata and C. capensis. The novel species appears to be geographically widely distributed, having been detected in Croatia, Spain and Georgia, USA. Although no information is available regarding the possible pathogenicity of the novel species for vertebrate hosts, R. hoogstraalii sp. nov. has a cytopathic effect in Vero, CCE3 and ISE6 cells. Sequence analyses of the 16S rRNA, 17 kDa, gltA, ompA and ompB genes indicated that even though R. hoogstraalii sp. nov. was closely related to Rickettsia felis, it represents a separate species within the spotted fever group. The type strain of R. hoogstraalii sp. nov. is strain Croatica T (5DSM 22243 T 5UTMB 00003 T ).