First records of Secretargas transgariepinus (Argasidae) in Libya and Jordan: corrections of collection records and detection of microorganisms.

The primarily bat-associated argasid tick, Secretargas transgariepinus (White, 1846), is a member of the Afrotropical and southern Palaearctic fauna. Probably because of its secretive life style, little is known about this species and records of its collection are scant. Based on morphological revisions of the available specimens, we report new Middle Eastern records for this tick species that had been misidentified as other bat-associated argasid taxa. These specimens are larvae from three localities, and represent the first records of S. transgariepinus from two countries: one larva from Sabratha (Libya) was collected from an unidentified bat species (possibly Eptesicus isabellinus), seven larvae from Azraq-Shishan (Jordan), and 78 larvae from Shamwari (Jordan) were all collected from Otonycteris hemprichii. Twenty larvae from Shamwari were also tested for the presence of both, viral or bacterial microorganisms by PCR. Three ticks were found to be infected with the Murid gammaherpesvirus 68 (MHV-68), one with Borrelia burgdorferi sensu lato, and four with a Rickettsia sp. closely related to Rickettsia slovaca. The findings represent a first evidence for the occurrence of these possible pathogens in S. transgariepinus.

Bartonella in bat flies from the Egyptian fruit bat in the Middle East.

In the family of fruit bats, Pteropodidae Gray, 1821, as in the third most diverse group of bats (Chiroptera), the bacterium of the genus Bartonella was detected in several species as well as in a few species of their insect ectoparasites in some tropical and sub-tropical regions of the Old World. The Egyptian fruit bat, Rousettus aegyptiacus (Geoffroy, 1810), is one of the most widespread fruit bats, occurring between South Africa, Senegal, and Pakistan. In this bat species, Candidatus Bartonella rousetti has been detected in three African populations in Nigeria, Kenya, and Zambia. This fruit bat, however, also occurs in the Palaearctic, an area isolating the species geographically and phylogenetically from the Afrotropical part of its distribution range. We screened the blood-sucking bat flies (family Nycteribiidae) from R. aegyptiacus for the presence of the Bartonella bacteria. A rich material of bat fly Eucampsipoda aegyptia (Macquart, 1850), a monoxenous ectoparasite of the Egyptian fruit bats, was collected at 26 localities in seven countries (Egypt, Iran, Jordan, Lebanon, Oman, United Arab Emirates, and Yemen) of the Middle East in 2007-2013. The DNA isolates from the bat flies were subjected to a three-marker (gltA, ssrA, and intergenic spacer region, ITS) multilocus sequence analysis. Based on the amplification of the fragment of ssrA gene by a real-time PCR, 65 E. aegyptia samples from 19 localities in all seven countries were positive for the bacteria. One to five Bartonella-positive individuals of E. aegyptia were collected per one individual of R. aegyptiacus. An analysis of the ITS and gltA genes indicated the presence of an uncultured Bartonella sp., belonging to the Cand. B. rousetti genogroup, identified from populations of the Egyptian fruit bat in Africa. These results support the hypothesis that Bartonella's diversity corresponds to its host's diversity (and phylogenetic structure). Specific lineages of pathogens are present in specific phylogenetic groups of bats.

Reticulinasus salahi (Acarina: Argasidae), a tick of bats and man in the Palaearctic and Afrotropics: review of records with the first pathogens detected.

The soft ticks of the genus Reticulinasus Schulze, 1941 (family Argasidae Koch, 1844) are ectoparasites of the fruit bats of the Old World (Pteropodidae). Reticulinasus salahi (Hoogstraal, 1953) is the only representative of this genus that occurs in the western part of the Palaearctic. This unusual distribution reflects the distributon range of its primary host, Rousettus aegyptiacus (Geoffroy, 1810). In this contribution, we present a revised review of records of this tick that were made in two periods, 1951-1966 (records from Egypt, Israel, Jordan, Spain) and 2005-2019 (Cyprus, Iran, Oman), and additionally, we present notes, re-determinations, new records, and summary of hosts of this tick. Besides the primary host, the revised list of hosts comprises two bats (Taphozous perforatus Geoffroy, 1818, Otonycteris hemprichii Peters, 1859) and the human (Homo sapiens Linnaeus, 1758). We also tried to identify pathogens in specimens of this tick collected from R. aegyptiacus in Oman. The DNA of the Mouse herpesvirus strain 68 (MHV-68), of two bacteria, Borellia burgdorferii sensu lato, and Ehrlichia sp. almost identical (98%) with Candidatus Ehrlichia shimanensis was detected in several larvae specimens.

Bartonella, Rickettsia, Babesia, and Hepatozoon Species in Fleas (Siphonaptera) Infesting Small Mammals of Slovakia (Central Europe).

Fleas (Siphonaptera) as obligate, blood-feeding ectoparasites are, together with ticks, hosted by small mammals and can transmit causative agents of serious infections. This study aimed to determine and characterize the presence and genetic diversity of Bartonella, Rickettsia, and apicomplexan parasites (Babesia, Hepatozoon) in fleas feeding on small mammals from three different habitat types (suburban, natural, and rural) in Slovakia. The most common pathogen in the examined fleas was Bartonella spp. (33.98%; 95% CI: 30.38-37.58), followed by Rickettsia spp. (19.1%; 95% CI: 16.25-22.24) and apicomplexan parasites (4.36%; 95% CI: 2.81-5.91). Bartonella strains belonging to B. taylorii, B. grahamii, B. elizabethae, Bartonella sp. wbs11, and B. rochalimae clades were identified in Ctenophthalmus agyrtes, C. congener, C. assimilis, C. sciurorum, C. solutus, C. bisoctodentatus, Palaeopsylla similis, Megabothris turbidus, and Nosopsyllus fasciatus within all habitats. The presence of Rickettsia helvetica, R. monacensis, and rickettsiae, belonging to the R. akari and R. felis clusters, and endosymbionts with a 96-100% identity with the Rickettsia endosymbiont of Nosopsyllus laeviceps laeviceps were also revealed in C. agyrtes, C. solutus, C. assimilis, C. congener, M. turbidus, and N. fasciatus. Babesia and Hepatozoon DNA was detected in the fleas from all habitat types. Hepatozoon sp. was detected in C. agyrtes, C. assimilis, and M. turbidus, while Babesia microti was identified from C. agyrtes, C. congener, and P. similis. The present study demonstrated the presence of zoonotic pathogens in fleas, parasitizing the wild-living small mammals of southwestern and central Slovakia and widens our knowledge of the ecology and genomic diversity of Bartonella, Rickettsia, Babesia, and Hepatozoon.

Ticks and their epidemiological role in Slovakia: from the past till present.

In Slovakia, 22 tick species have been found to occur to date. Among them, Ixodes ricinus, Dermacentor reticulatus, D. marginatus and marginally Haemaphysalis concinna, H. inermis and H. punctata have been identified as the species of public health relevance. Ticks in Slovakia were found to harbour and transmit zoonotic and/or potentially zoonotic agents such as tick-borne encephalitis virus (TBEV), spirochaetes of the Borrelia burgdorferi sensu lato (s.l.) complex, the relapsing fever sprirochaete Borrelia miyamotoi, bacteria belonging to the orders Rickettsiales (Rickettsia spp., Anaplasma phagocytophilum, Neoehrlichia mikurensis), Legionellales (Coxiella burnetii), and Thiotrichales (Francisella tularensis), and Babesia spp. parasites (order Piroplasmida). Ixodes ricinus is the principal vector of the largest variety of microorganisms including viruses, bacteria and piroplasms. TBEV, B. burgdorferi s.l., rickettsiae of the spotted fever group, C. burnetii and F. tularensis have been found to cause serious diseases in humans, whereas B. miyamotoi, A. phagocytophilum, N. mikurensis, Babesia microti, and B. venatorum pose lower or potential risk to humans. Distribution of TBEV has a focal character. During the last few decades, new tick-borne encephalitis (TBE) foci and their spread to new areas have been registered and TBE incidence rates have increased. Moreover, Slovakia reports the highest rates of alimentary TBE infections among the European countries. Lyme borreliosis (LB) spirochaetes are spread throughout the distribution range of I. ricinus. Incidence rates of LB have shown a slightly increasing trend since 2010. Only a few sporadic cases of human rickettsiosis, anaplasmosis and babesiosis have been confirmed thus far in Slovakia. The latest large outbreaks of Q fever and tularaemia were recorded in 1993 and 1967, respectively. Since then, a few human cases of Q fever have been reported almost each year. Changes in the epidemiological characteristics and clinical forms of tularaemia have been observed during the last few decades. Global changes and development of modern molecular tools led to the discovery and identification of emerging or new tick-borne microorganisms and symbionts with unknown zoonotic potential. In this review, we provide a historical overview of research on ticks and tick-borne pathogens in Slovakia with the most important milestones and recent findings, and outline future directions in the investigation of ticks as ectoparasites and vectors of zoonotic agents and in the study of tick-borne diseases.

Birds Belonging to the Family Paridae as Another Potential Reservoir of Murine Gammaherpesvirus 68.

Ecology and epidemiology of murine gammaherpesvirus 68 (MHV-68) have been intensively studied since the isolation of the virus from murid rodents in 1976. This virus was detected in various mammalian species that share the biotope with rodent reservoirs of MHV-68. However, a survey of MHV-68 in birds has not so far been performed. Therefore, the aim of this study was to investigate the presence of MHV-68 in blood samples from two bird species captured at four localities in Slovakia. Using the nested PCR targeting ORF50 gene of MHV-68, we confirmed the presence of MHV-68 DNA in 9 out of 57 blood samples from Great tits (Parus major) (prevalence 15.8%, confidence interval [95% CI]: 8.5-27.4) and in 3 out of 43 blood samples from Eurasian blue tits (Cyanistes caeruleus) (prevalence 7.0%, 95% CI: 2.4-18.6). Our results suggest that not only mammals but also birds may serve as reservoirs for MHV-68, providing further evidence that MHV-68 is capable of frequent cross-species transmission.

Pathogenic microorganisms in ticks removed from Slovakian residents over the years 2008-2018.

A total of 750 ticks feeding on humans were collected during the years 2008-2018. The majority of ticks (94.8 %) came from Slovakia, with 3.5 % from the Czech Republic, 0.9 % from Austria, and 0.3 % from Hungary. Travellers from Ukraine, Croatia, France, and Cuba also brought one tick from each of these countries. The majority of the analysed ticks were identified as Ixodes ricinus (94.3 %). Dermacentor reticulatus (0.93 %), Haemaphysalis concinna (0.1 %), Haemaphysalis sp. (0.1 %), Ixodes arboricola (0.1 %), and Rhipicephalus sp. (0.1 %) were also encountered. The most frequently found stage of I. ricinus was the nymph (69.9 %) followed by adult females (20.4 %) and larvae (8.3 %). Ticks were predominantly found on children younger than 10 years (46.3 %) and adults between 30-39 years (21.4 %). In children younger than 10 years, the ticks were usually found on the head, while in other age categories, the ticks were predominantly attached to legs. Ticks were further individually analysed for the presence of Rickettsia spp., Coxiella burnetii, Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Neoehrlichia mikurensis, Bartonella spp. and Babesia spp. The overall prevalences of tick-borne bacteria assessed in I. ricinus ticks acquired in Slovakia were: rickettsiae 25.0 % (95 % CI: 21.7-28.2), B. burgdorferi s.l. 20.5 % (95 % CI: 17.4-23.5), A. phagocytophilum 13.5 % (95 % CI: 10.9-16.0), Babesia spp. 5.2 % (95 % CI: 3.5-6.9), C. burnetii 3.0 % (95 % CI: 1.5-4.6), and N. mikurensis 4.4 % (95 % CI: 2.0-6.8). Pathogenic species Rickettsia raoultii, Rickettsia helvetica, Rickettsia monacensis, A. phagocytophilum, Borrelia garinii, Borrelia afzelii, Borrelia valaisiana, Babesia microti, and Babesia divergens were identified in D. reticulatus and I. ricinus ticks.

Circulation of Rickettsia species and rickettsial endosymbionts among small mammals and their ectoparasites in Eastern Slovakia.

Bacteria belonging to the genus Rickettsia are known as causative agents of vector-borne zoonotic diseases, such as spotted fevers, epidemic typhus and endemic typhus. Different species of ticks, mites and fleas could act as reservoirs and arthropod vectors of different pathogenic Rickettsia species. The aim of this work was to establish active surveillance of Rickettsia spp. in mites, ticks and fleas collected from small mammals (rodents and shrews) in Eastern Slovakia. A total of 964 animal ear biopsies, 871 mites, 667 ticks and 743 fleas were collected from small mammals in the Kosice region, Eastern Slovakia. All specimens were identified using specialized taxonomic keys, and were conserved in ethanol until DNA extraction was performed. After DNA extraction, identification of Rickettsia species was performed by PCR-based methods. The total prevalence of rickettsiae from ear biopsies was 4.6% (95% CI, 3.2-5.9), in tested mites 9.3% (95% CI, 7.4-11.2), 17.2% (95% CI, 14.3-20.1) in I. ricinus ticks and 3.5% (95% CI, 2.2-4.8) in fleas. Sequence analysis of the partial gltA gene and Rickettsia helvetica-, Rickettsia slovaca-, Rickettsia raoultii- species specific real-time PCR tests revealed the presence of R. helvetica, R. slovaca, unidentified Rickettsia and rickettsial endosymbionts. These pathogenic and symbiotic species were confirmed in the following ectoparasite species-Laelaps jettmari, Haemogamasus nidi, Laelaps agilis and Eulaelaps stabularis mites, Ixodes ricinus ticks, Ctenophthalmus solutus, C. assimilis and Megabothris turbidus fleas infesting host-Apodemus agrarius, A. flavicollis, Microtus arvalis and Myodes glareolus small mammals. These results confirm the circulation of R. helvetica, R. slovaca, unidentified Rickettsia and rickettsial endosymbionts in mites, ticks and fleas collected on small mammals in the Kosice region, Eastern Slovakia.

Comparative proteomics of the vector Dermacentor reticulatus revealed differentially regulated proteins associated with pathogen transmission in response to laboratory infection with Rickettsia slovaca.

BACKGROUND: Tick-borne rickettsial diseases are caused by pathogens acquired from hard ticks. In particular, Rickettsia slovaca, a zoonotic infectious bacterium causing tick-borne lymphadenopathy (TIBOLA), is transmitted by the vectors Dermacentor spp. that can be found all over Europe. Although recent studies point out the extreme complexity of bacteria-induced effects in these blood-feeding vectors, the knowledge of individual molecules involved in the preservation and transmission of the pathogen is still limited. System biology tools, including proteomics, may contribute greatly to the understanding of pathogen-tick-host interactions. METHODS: Herein, we performed a comparative proteomics study of the tick vector Dermacentor reticulatus that was experimentally infected with the endosymbiotic bacterium R. slovaca. Rickettsia-free ticks, collected in the southern region of Slovakia, were infected with the bacterium by a capillary tube-feeding system, and the dynamics of infection was assessed by quantitative PCR method after 5, 10, 15 and 27 days. RESULTS: At the stage of controlled proliferation (at 27 dpi), 33 (from 481 profiled) differentially abundant protein spots were detected on a two-dimensional gel. From the aforementioned protein spots, 21 were successfully identified by tandem mass spectrometry. CONCLUSIONS: Although a few discovered proteins were described as having structural or housekeeping functions, the vast majority of the affected proteins were suggested to be essential for tick attachment and feeding on the host, host immune system evasion and defensive response modulation to ensure successful pathogen transmission.

Simultaneous Occurrence of Borrelia miyamotoi, Borrelia burgdorferi Sensu Lato, Anaplasma phagocytophilum and Rickettsia helvetica in Ixodes ricinus Ticks in Urban Foci in Bratislava, Slovakia.

BACKGROUND: Questing Ixodes ricinus ticks were collected in two urban parks (Zelezná studienka and Horský park) of the capital city of Slovakia, Bratislava, during two consecutive years in 2011 and 2012. A total of 932 ticks were analyzed for the presence of tick-borne agents: B. miyamotoi, B. burgdorferi s.l., A. phagocytophilum and R. helvetica. RESULTS: PCR analysis confirmed the presence of all pathogens at both localities. The overall infection prevalence of B. miyamotoi, B. burgdorferi s.l., A. phagocytophilum and R. helvetica was 0.75, 13.2, 5.6 and 8.9%, respectively. B. burgdorferi s.l. positive samples were represented by six genospecies. The most frequent one was B. afzelii followed by B. garinii and B. valaisiana. CONCLUSION: Our study confirms the presence of I. ricinus ticks and at least nine tick-borne bacterial agents in city forest parks, which are used for recreational purposes. Ordination analysis revealed significant differences in the composition of pathogens with respect to study site location, time of season and ambient temperature, despite the fact that both sites are located relatively close to one another within the city.

Evaluation of the possible use of genus Mentha derived essential oils in the prevention of SENLAT syndrome caused by Rickettsia slovaca.

ETHNOPHARMACOLOGICAL RELEVANCE: Essential oils and essential oil bearing medicinal and culinary plants have a long tradition of being used to combat infection, treat various conditions, and promote and restore health. Mint oils are traditionally applied to repel insects and treat various conditions including wounds, skin infections, inflammation, eczema, urticaria, psoriasis, scabies and insect bites. They are among essential oils promoted as a natural way to prevent tick-borne diseases and recommended as ingredients in various homemade repellent mixtures and tick-bite treatments. AIM OF THE STUDY: The aim of this study was to evaluate the effect of three most common mint oils - peppermint (Mentha x piperita L.), cornmint (M. arvensis L.), and spearmint (M. spicata L.) on obligate intracellular tick-borne bacterium Rickettsia slovaca. MATERIALS AND METHODS: Influence of mint oils on R. slovaca replication in Vero cells initially infected by lower (106) or higher (108) number of rickettsial particles was tested during in vitro cultivation with daily change of medium. qPCR and RT-qPCR based growth curves and linear mixed effect models were applied to evaluate the growth inhibition. Peppermint oil was further tested in pilot in vivo study on experimentally infected ticks. RESULTS: Two of the tested essential oils, peppermint and cornmint, significantly inhibited rickettsial growth. On average, peppermint oil reduced the amount of rickettsiae present on day 4 post infection up to 0.05% of the rickettsial load present in the respective controls. Cornmint oil decreased the amount of rickettsiae to 0.09% of control. Peppermint oil also significantly reduced the number of living rickettsiae in artificially infected ticks. CONCLUSIONS: Present study showed that essential oils with antimicrobial properties may also inhibit tick-transmitted bacteria, and thus their possible use as preventative measures against tick-borne diseases is worth further research.

Low-cost light-induced therapy to treat rickettsial infection.

Antimicrobial photodynamic therapy uses a nontoxic photosensitizer with the assistance of harmless visible light to activate the photosensitizer. Consequently, the excited state of the photosensitizer interacts with molecular oxygen to produce reactive oxygen species, which have the antimicrobial effect. In this study, we evaluated the effect of photodynamic therapy on Vero cells infected with rickettsia using methylene blue as a photosensitizer along with red light. A significant reduction (by 96%) in the number of viable Rickettsia slovaca was determined by quantitative RT-PCR 48 h after the treatment with methylene blue followed by 30 min of red light excitation. A statistically significant reduction of R. slovaca was also recorded with pretreatment (by 99%). To the best of our knowledge, this result is the first one in the literature to confirm the effectiveness of photodynamic therapy for the elimination of R. slovaca and to suggest this technique as a good supportive treatment for rickettsial infections.

Diverse tick-borne microorganisms identified in free-living ungulates in Slovakia.

BACKGROUND: Free-living ungulates are hosts of ixodid ticks and reservoirs of tick-borne microorganisms in central Europe and many regions around the world. Tissue samples and engorged ticks were obtained from roe deer, red deer, fallow deer, mouflon, and wild boar hunted in deciduous forests of south-western Slovakia. DNA isolated from these samples was screened for the presence of tick-borne microorganisms by PCR-based methods. RESULTS: Ticks were found to infest all examined ungulate species. The principal infesting tick was Ixodes ricinus, identified on 90.4% of wildlife, and included all developmental stages. Larvae and nymphs of Haemaphysalis concinna were feeding on 9.6% of wildlife. Two specimens of Dermacentor reticulatus were also identified. Ungulates were positive for A. phagocytophilum and Theileria spp. Anaplasma phagocytophilum was found to infect 96.1% of cervids, 88.9% of mouflon, and 28.2% of wild boar, whereas Theileria spp. was detected only in cervids (94.6%). Importantly, a high rate of cervids (89%) showed mixed infections with both these microorganisms. In addition to A. phagocytophilum and Theileria spp., Rickettsia helvetica, R. monacensis, unidentified Rickettsia sp., Coxiella burnetii, "Candidatus Neoehrlichia mikurensis", Borrelia burgdorferi (s.l.) and Babesia venatorum were identified in engorged I. ricinus. Furthermore, A. phagocytophilum, Babesia spp. and Theileria spp. were detected in engorged H. concinna. Analysis of 16S rRNA and groEL gene sequences revealed the presence of five and two A. phagocytophilum variants, respectively, among which sequences identified in wild boar showed identity to the sequence of the causative agent of human granulocytic anaplasmosis (HGA). Phylogenetic analysis of Theileria 18S rRNA gene sequences amplified from cervids and engorged I. ricinus ticks segregated jointly with sequences of T. capreoli isolates into a moderately supported monophyletic clade. CONCLUSIONS: The findings indicate that free-living ungulates are reservoirs for A. phagocytophilum and Theileria spp. and engorged ixodid ticks attached to ungulates are good sentinels for the presence of agents of public and veterinary concern. Further analyses of the A. phagocytophilum genetic variants and Theileria species and their associations with vector ticks and free-living ungulates are required.

Seasonal Patterns in the Prevalence and Diversity of Tick-Borne Borrelia burgdorferi Sensu Lato, Anaplasma phagocytophilum and Rickettsia spp. in an Urban Temperate Forest in South Western Slovakia.

In Europe, Ixodes ricinus is the most important vector of tick-borne zoonotic bacteria. It transmits spirochaetes from the Borrelia burgdorferi sensu lato complex, Anaplasma phagocytophilum and Rickettsia spp. Although spatial differences in the prevalence of tick-borne pathogens have been intensively studied, seasonal (within-year) fluctuations in the prevalence of these pathogens within sites are often overlooked. We analyzed the occurrence and seasonal dynamics of Ixodes ricinus in an urban forest in Bratislava, Slovakia. Furthemore, we examined temporal trends in the community structure of B. burgdorferi s.l., A. phagocytophilum and Rickettsia spp. in questing and bird-feeding ticks. The total prevalence for B. burgdorferi s.l. in questing I. ricinus was 6.8%, involving six genospecies with the dominance of bird-associated B. garinii and B. valaisiana.A. phagocytophilum, R. helvetica and R. monacensis occurred in 5.9%, 5.0% and 0.2% of questing ticks, respectively. In total, 12.5% and 4.4% of bird-feeding I. ricinus ticks carried B. burgdorferi s.l. and R. helvetica. The total prevalence of B. burgdorferi s.l. in our study site was two times lower than the mean prevalence for Europe. In contrast, A. phagocytophilum prevalence was significantly higher compared to those in other habitats of Slovakia. Our results imply that tick propagation and the transmission, suppression and seasonal dynamics of tick-borne pathogens at the study site were primarily shaped by abundance and temporal population fluctuations in ruminant and bird hosts.

Diversity of Coxiella-like and Francisella-like endosymbionts, and Rickettsia spp., Coxiella burnetii as pathogens in the tick populations of Slovakia, Central Europe.

Ticks are important vectors of pathogens affecting humans and animals worldwide. They do not only carry pathogens but diverse commensal and symbiotic microorganisms are also present in ticks. A molecular screening for tick-borne pathogens and endosymbionts was carried out in Ixodes ricinus, Dermacentor reticulatus and Haemaphysalis inermis questing ticks collected in Slovakia. The presence of Rickettsia spp., Coxiella burnetii, Coxiella-like and Francisella-like microorganisms was evaluated by PCR in 605 individuals and by randomly sequencing 66 samples. Four species of rickettsiae (R. raoultii, R. slovaca, R. helvetica and R. monacensis) were identified and reported with an overall prevalence range between 0.4 and 50.3% (±8.0) depending on tick species, sex and locality. Partial sequencing of the gltA gene of 5 chosen samples in H. inermis showed 99% identity with Candidatus Rickettsia hungarica. The total prevalence of C. burnetii in ticks was 2.2 ±â€¯1.7%; bacteria were confirmed in I. ricinus and D. reticulatus ticks. The sequences from 2 D. reticulatus males and 1 I. ricinus female ticks were compared to GenBank submissions and a 99.8% match was obtained with the pathogenic C. burnetii. Coxiella-like endosymbionts were registered in all three species of ticks from all studied sites with an average prevalence of 32.7 ±â€¯3.7%. A phylogenetic analysis of this Coxiella sp. showed that it does not group with the pathogenic C. burnetii. The prevalence of Francisella-like microorganisms in questing ticks was 47.9 ±â€¯3.9%, however H. inermis (n = 108) were not infested. Obtained sequences were 98% identical with previously identified Francisella-like endosymbionts in D. reticulatus and I. ricinus. Coxiella-like and Francisella-like microorganisms were identified for the first time in Slovakia, they might be considered as a non-pathogenic endosymbiont of I. ricinus, D. reticulatus and H. inermis, and future investigations could aim to assess their role in these ticks. However, this work provided further data and broadened our knowledge on bacterial pathogens and endosymbionts present in ticks in Slovakia to help understanding co-infestations, combined treatments and public health issues linked to tick bites.

Effect of Climate and Land Use on the Spatio-Temporal Variability of Tick-Borne Bacteria in Europe.

The incidence of tick-borne diseases caused by Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Rickettsia spp. has been rising in Europe in recent decades. Early pre-assessment of acarological hazard still represents a complex challenge. The aim of this study was to model Ixodes ricinus questing nymph density and its infection rate with B. burgdorferi s.l., A. phagocytophilum and Rickettsia spp. in five European countries (Italy, Germany, Czech Republic, Slovakia, Hungary) in various land cover types differing in use and anthropisation (agricultural, urban and natural) with climatic and environmental factors (Normalized Difference Vegetation Index (NDVI), Normalized Difference Water Index (NDWI), Land Surface Temperature (LST) and precipitation). We show that the relative abundance of questing nymphs was significantly associated with climatic conditions, such as higher values of NDVI recorded in the sampling period, while no differences were observed among land use categories. However, the density of infected nymphs (DIN) also depended on the pathogen considered and land use. These results contribute to a better understanding of the variation in acarological hazard for Ixodes ricinus transmitted pathogens in Central Europe and provide the basis for more focused ecological studies aimed at assessing the effect of land use in different sites on tick-host pathogens interaction.

Tick-borne pathogens and their reservoir hosts in northern Italy.

The aim of this study was to determine the occurrence of Anaplasma phagocytophilum, Rickettsia spp., Babesia spp., and Candidatus Neoehrlichia mikurensis in Ixodes spp. ticks removed from wildlife, domestic animals and humans in the Province of Trento (northern Italy) in order to better understand their ecology and provide public health professionals with an updated list of pathogens which should be considered during their diagnostic procedures after a tick bite. During 2011-2012, 848 feeding ticks at all life stages (adults, nymphs and larvae) from various hosts (wild ungulates, birds and rodents; domestic sheep, dogs and humans) were collected. The highest prevalences of A. phagocytophilum and Rickettsia spp. were detected in adult and nymphal tick stages feeding on wild ungulates (11.4% prevalence for both pathogens), while the Babesia spp. prevailed in nymphal and larval ticks feeding on wild birds (7.7%). A wide spectrum of tick-borne agents was present in larval ticks: those detached from wild ungulates were positive for A. phagocytophilum, B. venatorum, R. helvetica, R. monacensis and R. raoultii, while those removed from wild rodents were positive for B. venatorum, R. helvetica, R. monacensis and Ca. N. mikurensis, and ticks from wild birds carried A. phagocytophilum, B. venatorum, B. capreoli and R. helvetica. This study provides evidence of circulation of five tick-borne pathogens not reported in this region before, specifically R. raoultii, R. monacensis, B. venatorum, B. capreoli and B. microti. Furthermore, it discusses the epidemiological role of the animal species from which the ticks were collected highlighting the needs for more experimental studies especially for those pathogens where transovarial transmission in ticks has been demonstrated.

Diversity and prevalence of Bartonella species in small mammals from Slovakia, Central Europe.

Wild-living rodents are important hosts for zoonotic pathogens. Bartonella infections are widespread in rodents; however, in Slovakia, knowledge on the prevalence of these bacteria in small mammals is limited. We investigated the prevalence and diversity of Bartonella species in the spleens of 640 rodents of six species (Apodemus flavicollis, Apodemus sylvaticus, Myodes glareolus, Microtus arvalis, Microtus subterraneus, and Micromys minutus) and in the European mole (Talpa europaea) from three different habitat types in south-western and central Slovakia. Overall, the prevalence of Bartonella spp. in rodents was 64.8%; a rate of 73.8% was found in natural habitat (deciduous forest), 56.0% in suburban forest park and 64.9% in rural habitat. Bartonella spp. were detected in 63.0% of A. flavicollis, 69% of My. glareolus and 61.1% of M. arvalis and in T. europaea. However, Bartonella were not found in the other examined rodents. Molecular analyses of the 16S-23S rRNA intergenic spacer region revealed the presence of four different Bartonella spp. clusters. We identified B. taylorii, B. rochalimae, B. elizabethae, B. grahamii and Bartonella sp. wbs11 in A. flavicollis and My. glareolus. Bartonella genotypes ascribed to B. taylorii and B. rochalimae were found in M. arvalis. B. taylorii was identified in T. europaea. Questing Ixodes ricinus ticks that were collected at the study sites were not infected with Bartonella. This study improves our understanding of the ecoepidemiology of Bartonella spp. in Europe and underlines the necessity for further research on Bartonella-host-vector associations and their consequences on animal and human health in Slovakia.

The repellent efficacy of eleven essential oils against adult Dermacentor reticulatus ticks.

Dermacentor reticulatus ticks are among the most important arthropod vectors of zoonotic disease agents in Europe. Eleven essential oils, namely basil (Ocimum basilicum), bergamot (Citrus bergamia), clove bud (Syzygium aromaticum), citronella (Cymbopogon winterianus), creeping thyme (Thymus serpyllum), lavender (Lavandula angustifolia), lemon-scented gum (Corymbia citriodora), marjoram (Origanum majorana), peppermint (Mentha piperita), spearmint (M. spicata), and red thyme (Thymus vulgaris) were tested for repellency against adult D. reticulatus ticks at concentrations of 1 and 3%. Clove bud, creeping thyme and red thyme essential oils were the most efficient - repelling 83, 82 and 68% of ticks when diluted to 3%, respectively. The mixture of creeping thyme and citronella containing 1.5% of each showed higher repellency (91%) than individual essential oils at the concentration of 3%.