Despite being a small country, Kosovo represents one of the few foci of Crimean-Congo hemorrhagic fever (CCHF) in Europe. The distribution of Kosovar tick vectors and the evolution of CCHF virus in ticks are both as yet unknown. A better description of the extent and the genetic diversity of CCHFV in ticks from endemic settings is essential, in order to be controlled. We investigated the 2012 distribution of Kosovar ticks alongside the prevalence and the phylogeography of tick-derived CCHFV. Hyalomma marginatum dominated in the endemic municipalities with 90.2% versus 24.3% in the non-endemic regions. Of 1,102 tested ticks, 40 (3.6%) were CCHFV-positive, belonging to H. marginatum (29), Rhipicephalus bursa (10), and Ixodes ricinus (1). The virus strains clustered with clade V and VI related sequences. They fell into two lineages: Kosovo I and II. Kosovo I comprised strains recovered exclusively from R. bursa ticks and was closely related to AP92 prototype strain. Kosovo II clustered into Kosovo IIa, including human-derived strains, and IIb including only strains detected in H. marginatum and I. ricinus. Our phylogeographic reconstruction suggests two temporally distinct CCHFV introductions: the most probable location of the most recent common ancestor of Kosovo I lineage was in Greece (63 years ago) and that of lineages IIa-b in Turkey (35 years ago). After each CCHFV introduction into Kosovo, subsequent lineage expansions suggest periods of in situ evolution. The study provides the first insight into the genetic variability and the origin of CCHFV in ticks from Kosovo. Our findings indicate the spreading of CCHFV to non-endemic areas, which underlines the importance of further studies in order to monitor and predict future CCHF outbreaks in Kosovo. The AP92-like strains appear to be more widespread than previously thought and may provide a promising target for experimental studies due to their assumed low pathogenicity. Crimean-Congo hemorrhagic fever virus (CCHFV) is the most widespread tick-borne virus affecting humans. Its distribution coincides with that of Old World Hyalomma tick vectors. The virus circulates in an enzootic cycle, where hard ticks are both vectors and reservoirs, and mammals the amplifying hosts. Accidentally, humans become infected as dead-end hosts. One of the few CCHF-endemic European countries is Kosovo, where an annual average of ten sporadic cases, with a mean mortality rate of 18%, are reported. In this study, the tick distribution, CCHFV prevalence and the phylogeography of the tick-derived strains from 2012 were investigated. The majority of ticks belonged to Hyalomma marginatum, the main vector of CCHFV. Indeed, of 40 CCHFV-positive ticks, 29 were H. marginatum. The phylogenetic analyses revealed that 30 viral strains formed a distinct lineage within clade V (Europe 1) and were closely related to previously known strains from humans and ticks from the Balkans. However, ten isolates from Rhipicephalus bursa ticks, collected in a non-endemic region, turned out to cluster within a very divergent CCHFV clade (Europe 2), so far only known from Greece and Turkey. Since these strains presumably develop only subclinical or mild infections in humans, they might be suitable for experimental vaccine studies.

Background: Vector-borne diseases (VBD) are of growing global importance. Sand flies are potential vectors for phleboviruses (family Phenuiviridae) including Toscana virus (TOSV), Sicilian virus, Sandfly fever, Naples virus, and Leishmania parasites in Europe. To date, only two phlebotomine species have been recorded for Germany: Phlebotomus perniciosus and Phlebotomus mascittii. This study updates the distribution and abundance of the two occurring species. Methods: An entomological field study was carried out during 2015–2018 to assess the abundance of sand flies in Southwest Germany within the federal states Baden-Wuerttemberg (BW) and Rhineland-Palatinate (RLP). A total of 176 collection sites were examined using CDC light traps. Results: A total of 149 individuals of P. mascittii were collected. During 2015–2018, P. mascittii was found at all sites known positive from previous studies and was detected at 15 additional sites previously unknown for the presence of sand flies. Although the environment has changed considerably in 30 years, no significant difference in sand fly dynamics and distribution was found. Phlebotomus perniciosus has only been trapped once since 2001. Conclusions: This study showed that sand flies occur in different areas in Southern Germany where they had not been recorded previously. Therefore, it can be assumed that they are more widespread than expected. In addition, sand flies could be found over several years at the same trapping sites, indicating population stability. This supports the need for continued surveillance of possible vector populations and urgent clarification of the vector competence of P. mascittii.

Background: Knowledge of the potential vector role of Culicidae mosquitoes in Germany is very scanty, and until recently it was generally assumed that they are not involved in the transmission of anthroponotic or zoonotic pathogens in this country. However, anticipated changes in the course of global warming and globalization may alter their status.Methods: We conducted a molecular mass screening of mosquitoes for filarial parasites using mitochondrial 12S rRNA-based real-time PCR.Results: No parasites causing disease in humans such asDirofilariaspp. were detected in about 83,000 mosquitoes tested, which had been collected in 2009 and 2010 in 16 locations throughout Germany. However, minimum infection rates of up to 24 per 1000 mosquitoes were revealed, which could be attributed to mosquito infection withSetaria tundraand a yet unidentified second parasite.Setaria tundrawas found to be widespread in southern Germany in various mosquito species, exceptCulexspp. In contrast, the unidentified filarial species was exclusively found inCulexspp. in northern Baden-Württemberg, and is likely to be a bird parasite.Conclusions: Although dirofilariasis appears to be emerging and spreading in Europe, the absence ofDirofilariaspp. or other zoonotic filariae in our sample allows the conclusion that the risk of autochthonous infection in Germany is still very low. Potential vectors ofS. tundrain Germany areOchlerotatus sticticus,Oc. cantans,Aedes vexansandAnopheles claviger. Technically, the synergism between entomologists, virologists and parasitologists, combined with state-of-the-art methods allows a very efficient near-real-time monitoring of a wide spectrum of both human and veterinary pathogens, including new distribution records of parasite species and the incrimination of their potential vectors.