The biodiversity of Culicoides from eastern Slovakia was investigated by light trapping. An integrative taxonomy approach combining DNA barcode sequence and morphological analyses was used to accurately identify specimens. Five species were newly recorded from Slovakia: Culicoides picturatus Kremer & Deduit, C. gejgelensis Dzhafarov, C. clastrieri Callot et al., C. griseidorsum Kieffer and C. odiatus Austen. The checklist of the Culicoides species recorded from SK has been updated to 63 species and barcode sequence data is provided for 8 species not previously available on GenBank. Conflict between results from molecular and morphological analyses resulted in the discovery of some potentially new cryptic species and the inability of DNA barcodes to distinguish C. festivipennis Kieffer from C. clastrieri, C. salinarius Kieffer from C. manchuriensis Tokunaga and C. pallidicornis Kieffer from C. subfasciipennis Kieffer. These conflicts suggest further study is required to clarify the status of these species.

Species of the genus Culicoides Latreille, 1809 (Diptera: Ceratopogonidae) are mainly known as vectors of arboviruses such as bluetongue (BTV) and Schmallenberg (SBV). Among the known vectors, few species within the subgenus Culicoides Latreille, 1809 have been implicated in the transmission of BTV and SBV. Nevertheless, phylogenetic studies had revealed the presence of cryptic and undescribed species in Europe, raising questions about their vectorial role. A previous integrative study, associating morphology and barcode data, raised the hypothesis of the presence of undescribed species in Slovakia. The present study, combining morphological and molecular approaches, is aimed to support the hypothesis and a description of Culicoides bysta n. sp. is provided. Series of male and female specimens were dissected and several of them were sequenced for the barcode region of the mitochondrial cytochrome c oxidase subunit 1 gene (cox1). Bayesian inference phylogenetic analyses based on 72 cox1 sequences of the species belonging to the Pulicaris group of the subgenus Culicoides, were carried out and the frequencies of intra/interspecific variations were analyzed. The morphology of abundant material of the new species (31 females and 12 males) was examined and compared with the paratypes of Culicoides boyi Nielsen, Kristensen & Pape, 2015 and with specimens of Culicoides pulicaris Linnaeus, 1758. For females, suture distances on the eyes were newly evaluated as a diagnostic character and for males we assessed a new measurement on the ninth tergite and on the apicolateral processes. Both phylogenetic analysis and barcode distances supported the distinct status of the new species, Culicoides bysta n. sp. described as a member of the Pulicaris group based on the morphology of males and females. The new species is closely related to C. boyi and C. pulicaris but can be distinguished on the basis of the wing pattern and the ratio between the two eye sutures. Both newly evaluated characters, i.e. eyes in females and male genitalia appeared to be diagnostic for distinguishing the new species described herein. The vector potential of the recently described species C. boyi and C. bysta n. sp. to transmit arboviruses, such as BTV and SBV, is unknown. When considering these two species as being close to C. pulicaris, the previous data, such as the vector implication for C. pulicaris in BTV transmission, should be revaluated in future.

Biting midges of the genus Culicoides are vectors of important pathogens affecting domestic and wild animals and have played a major role in the re-emergence of new outbreaks of bluetongue (BTV) and Schmallenberg (SBV) viruses in Europe. To determine vector-host specificity, trophic preference from blood meal analysis is of major importance in the surveillance of arthropod-borne diseases. Of 28,752 specimens collected, we identified 17 Culicoides species and investigated a total of 48 host sequences from the blood meals. Culicoides obsoletus/C. scoticus, C. dewulfi, C. pulicaris, C. lupicaris, C. punctatus, C. newsteadi, C. riethi, and C. furcillatus were found to feed on mammals (cattle, horses, and humans), birds (domestic chickens), small rodents (Apodemus flavicollis), and hares (Lepus europaeus). To our knowledge, this is the first study investigating trophic preferences of Culicoides spp. in Slovakia. This study demonstrated that Culicoides species are able to feed on domesticated host vertebrates as well as birds, rodents, and humans.

This is the third in a planned series of data papers presenting modelled vector distributions produced during the ECDC and EFSA funded VectorNet project. The data package presented here includes those Culicoides vectors species first modelled in 2015 as part of the VectorNet gap analysis work namely C. imicola, C. obsoletus, C. scoticus, C. dewulfi, C. chiopterus, C. pulicaris, C. lupicaris, C. punctatus, and C. newsteadi. The known distributions of these species within the Project area (Europe, the Mediterranean Basin, North Africa, and Eurasia) are currently incomplete to a greater or lesser degree. The models are designed to fill the gaps with predicted distributions, to provide a) first indication of vector species distributions across the project geographical extent, and b) assistance in targeting surveys to collect distribution data for those areas with no field validated information. The models are based on input data from light trap surveillance of adult Culicoides across continental Europe and surrounding regions (71.8°N –33.5°S, – 11.2°W – 62°E), concentrated in Western countries, supplemented by transect samples in eastern and northern Europe. Data from central EU are relatively sparse. Funding statement: This work was carried out with support from the VectorNet framework contract OC/EFSA/AHAW/2013/02-FWC1 funded by the European Centre for Disease prevention and Control (ECDC) and the European Food Safety Agency (EFSA) and the PALE-Blu H2020 Project ID: 727393.

This epizootiological study was carried out to investigate the occurrence of canine babesiosis in southwestern Slovakia. The study focused on the proportion of the species of ticks serving as babesia vectors of babesiosis in the ticks collected from selected locations close to Komárno, in southwestern Slovakia. Additionally, observations were made on the health and overall clinical signs in dogs suspected of having babesiosis. In 2014 we collected ticks from vegetation by the drag cloth (flagging) method and also directly from dogs. A totally of 622 ticks were collected: 491 Dermacentor reticulatus and 131 Ix-odes ricinus. Representative samples of ticks (n = 103) were examined by the molecular method and the presence of Babesia spp. DNA was identified in 12.5 % of the Ixodes ricinus ticks collected by flagging and in 9.5 % of the Ixodes ricinus ticks collected from the dogs. Babesia canis (KU681325) with 90 bp sequence, 100 % identical with Babesia canis isolates from dogs for example from: Turkey (KF499115), Rumania (HQ662634), Croatia (FJ209025), Poland (EU622792) and Russia (AY962186), was confirmed after sequencing in one Ixodes ricinus female obtained from a dog. This was the first confirmation of the occurrence of B. canis in Ixodes ricinus ticks in Slovakia. In 2.2 % of the Dermacentor reticulatus ticks obtained from vegetation by flagging, we were able to diagnose the DNA of Babesia canis. In 4.8 % of the Dermacentor reticulatus ticks collected from dogs, the presence of Babesia spp. was confirmed. Thirty three dogs with suspicion of babesiosis were observed in an ambulance by their health and clinical signs. The loss of appetite was observed in 22 patients (66.7 %), apathy in 19 cases (57.6 %), and fever in 19 cases. Closer specification indicating babesiosis was finding engorged ticks on the dog bodies which occurred in 21 cases (63.6 %), haematuria in 8 cases (24.2 %), anaemia in 4 cases (12.1 %), and tremor in 6 cases (18.2 %). Other non-specific signs, such as diarrhoea, vomitus, slowed-down movement, ataxia and lacrimation were also observed in less than 18 % of the examined dogs. On the basis of the specific clinical signs, blood was withdrawn from 33 dogs for preparation of blood smears and in 19 of them (57.6 %) babesia in erythrocytes were confirmed microscopically.