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BackgroundDespite the known circulation of West Nile virus (WNV) and Usutu virus (USUV) in Slovakia, no formal entomological surveillance programme has been established there thus far.AimTo conduct contemporaneous surveillance of WNV and USUV in different areas of Slovakia and to assess the geographical spread of these viruses through mosquito vectors. The first autochthonous human WNV infection in the country is also described.MethodsMosquitoes were trapped in four Slovak territorial units in 2018 and 2019. Species were characterised morphologically and mosquito pools screened for WNV and USUV by real-time reverse-transcription PCRs. In pools with any of the two viruses detected, presence of pipiens complex group mosquitoes was verified using molecular approaches.ResultsAltogether, 421 pools containing in total 4,508 mosquitoes were screened. Three pools tested positive for WNV and 16 for USUV. USUV was more prevalent than WNV, with a broader spectrum of vectors and was detected over a longer period (June-October vs August for WNV). The main vectors of both viruses were Culex pipiens sensu lato. Importantly, WNV and USUV were identified in a highly urbanised area of Bratislava city, Slovakias' capital city. Moreover, in early September 2019, a patient, who had been bitten by mosquitoes in south-western Slovakia and who had not travelled abroad, was laboratory-confirmed with WNV infection.ConclusionThe entomological survey results and case report increase current understanding of the WNV and USUV situation in Slovakia. They underline the importance of vector surveillance to assess public health risks posed by these viruses.
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Culex , Culicidae , Flavivirus , Febre do Nilo Ocidental , Vírus do Nilo Ocidental , Animais , Flavivirus/genética , Humanos , Eslováquia/epidemiologia , Febre do Nilo Ocidental/diagnóstico , Febre do Nilo Ocidental/epidemiologia , Vírus do Nilo Ocidental/genéticaRESUMO
Ticks will diminish productivity among farm animals and transmit zoonotic diseases. We conducted a study to identify tick species infesting slaughter bulls from Adama City and to screen them for tick-borne pathogens. In 2016, 291 ticks were collected from 37 bulls in Adama, which were ready for slaughter. Ticks were identified morphologically. Total genomic DNA was extracted from ticks and used to test for Rickettsia spp. with real-time PCR. Species identification was done by phylogenetic analysis using sequencing that targeted the 23S-5S intergenic spacer region and ompA genes. Four tick species from two genera, Amblyomma and Rhipicephalus, were identified. Amblyomma cohaerens was the dominant species (n = 241, 82.8%), followed by Amblyomma variegatum (n = 22, 7.5%), Rhipicephalus pulchellus (n = 19, 6.5%), and Rhipicephalus decoloratus (n = 9, 3.0%). Among all ticks, 32 (11%) were positive for Rickettsia spp. and 15 (5.2%) of these were identified as R. africae comprising at least two genetic clades, occurring in A. variegatum (n = 10) and A. cohaerens (n = 5). The remainder of Rickettsia-positive samples could not be amplified due to low DNA yield. Furthermore, another 15 (5.2%) samples carried other pathogenic bacteria: Ehrlichia ruminantium (n = 9; 3.1%) in A. cohaerens, Ehrlichia sp. (n = 3; 1%) in Rh. pulchellus and A. cohaerens, Anaplasma sp. (n = 1; 0.5%) in A. cohaerens, and Neoehrlichia mikurensis (n = 2; 0.7%) in A. cohaerens. All ticks were negative for Bartonella spp., Babesia spp., Theileria spp., and Hepatozoon spp. We reported for the first time E. ruminatium, N. mikurensis, Ehrlichia sp., and Anaplasma sp. in A. cohaerens. Medically and veterinarily important pathogens were mostly detected from A. variegatum and A. cohaerens. These data are relevant for a One-health approach for monitoring and prevention of tick-borne disease transmission.
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Doenças dos Bovinos , Ixodidae , Rickettsia , Infestações por Carrapato , Doenças Transmitidas por Carrapatos , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Etiópia/epidemiologia , Masculino , Filogenia , Rickettsia/genética , Infestações por Carrapato/epidemiologia , Infestações por Carrapato/veterinária , Doenças Transmitidas por Carrapatos/epidemiologia , Doenças Transmitidas por Carrapatos/veterináriaRESUMO
Ticks are ectoparasites of a wide range of animals and are important vectors of numerous pathogens affecting humans, livestock, and pets. This study investigates possible correlations between selected factors, altitude, soil pH, and a factor called 'amount' (number of ticks examined in pooled samples) on the occurrence of I. ricinus ticks positive for selected tick-borne microorganisms. Questing I. ricinus ticks were collected in 2016 and 2017 across various altitudes, at two mountain ranges in central Slovakia. Tick pools were screened for the presence of Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato (Bbsl), Babesia/Theileria spp., Rickettsia spp., and tick-borne encephalitis virus (TBEV) using molecular methods. Regression analysis was employed to evaluate relationships between selected factors and the occurrence of vector-borne microorganisms. This study revealed a statistically significant influence of altitude on the occurrence of A. phagocytophilum; increasing altitude of the sampling site was associated with increased probability of pathogen occurrence. For Babesia/Theileria spp., neither altitude nor soil pH significantly affected pathogen occurrence. The occurrence of Bbsl was notably impacted by both altitude and soil pH; higher altitudes were associated with a decreased probability of pathogen presence, whereas higher soil pH increased the likelihood of pathogen occurrence. The presence of Rickettsia in a pooled sample was not affected by altitude and soil pH, but the 'amount' factor was a significant predictor, increasing the probability of pathogen detection. Neither altitude nor soil pH had a significant impact on TBEV occurrence. The regression models showed moderate goodness-of-fit levels to the data, underscoring their utility in examining the role of altitude and soil pH on pathogen occurrence. However, they explained only a small portion of the overall variance in pathogen occurrence, indicating the presence of other significant factors not covered in this study.
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Ticks are involved in the transmission a plethora of pathogens. To effectively control ticks and mitigate the risks associated with tick-borne diseases, it is important to implement tick control measures. These may include the use of acaricides as well as the development and implementation of an alternative, environmentally friendly tick management program that include practices such as habitat modification or establishing biological control. Ixodiphagus hookeri Howard is a tick-specific parasitoid wasp that predates on several species of ixodid ticks and could contribute to the control of the tick population. This work aimed to detect the presence of parasitoid wasps in ticks (Ixodidae) using genetic approaches. Several tick species of the genera Ixodes, Haemaphysalis, and Dermacentor, with a sympatric occurrence in the Slovak Karst National Park in southeastern Slovakia, were screened for the presence of wasps of the genus Ixodiphagus. The DNA of the parasitoids was detected in four tick species from three genera. This work presents the first molecular detection of parasitoids in two Dermacentor tick species, as well as the first molecular identification of Ixodiphagus wasps in Ixodes ricinus and Haemaphysalis concinna ticks from the Karst area. In the given area, it was observed that I. ricinus and H. concinna ticks are hyper-parasitized by wasps. Moreover, it was observed that wasps here can parasitize several tick species, some of which are of less significance for human and animal health (as they transmit fewer pathogens).
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In order to obtain a better understanding of tick-borne encephalitis virus (TBEV) strain movements in central Europe the E gene sequences of 102 TBEV strains collected from 1953 to 2011 at 38 sites in the Czech Republic, Slovakia, Austria and Germany were determined. Bayesian analysis suggests a 350-year history of evolution and spread in central Europe of two main lineages, A and B. In contrast to the east to west spread at the Eurasian continent level, local central European spreading patterns suggest historic west to east spread followed by more recent east to west spread. The phylogenetic and network analyses indicate TBEV ingressions from the Czech Republic and Slovakia into Germany via landscape features (Danube river system), biogenic factors (birds, red deer) and anthropogenic factors. The identification of endemic foci showing local genetic diversity is of paramount importance to the field as these will be a prerequisite for in-depth analysis of focal TBEV maintenance and long-distance TBEV spread.
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Vírus da Encefalite Transmitidos por Carrapatos/classificação , Vírus da Encefalite Transmitidos por Carrapatos/genética , Filogeografia , Proteínas do Envelope Viral/genética , Animais , Análise por Conglomerados , Vírus da Encefalite Transmitidos por Carrapatos/isolamento & purificação , Europa (Continente) , Genótipo , Mamíferos/virologia , Epidemiologia Molecular , Dados de Sequência Molecular , Análise de Sequência de DNA , Carrapatos/virologiaRESUMO
The distribution and prevalence of zoonotic pathogens infecting ixodid ticks in Western Europe have been extensively examined. However, data on ticks and tick-borne pathogens in Eastern Europe, particularly Ukraine are scarce. The objective of the current study was, therefore, to investigate the prevalence of Anaplasma phagocytophilum, Anaplasmataceae, Rickettsia spp., Babesia spp., Bartonella spp., and Borrelia burgdorferi sensu lato in engorged and questing ixodid ticks collected from five administrative regions (oblasts) of Ukraine, namely Chernivtsi, Khmelnytskyi, Kyiv, Ternopil, and Vinnytsia. The ticks were collected from both wild and domestic animals and from vegetation. Of 524 ixodid ticks collected, 3, 99, and 422 ticks were identified as Ixodes hexagonus, Ixodes ricinus, and Dermacentor reticulatus, respectively. DNA samples individually extracted from 168 questing and 354 engorged adult ticks were subjected to pathogen-specific PCR analyses. The mean prevalence in I. ricinus and D. reticulatus were, respectively: 10 % (10/97) and 3 % (12/422) for A. phagocytophilum; 69 % (67/97) and 52 % (220/422) for members of the Anaplasmataceae family; 25 % (24/97) and 28 % (117/422) for Rickettsia spp.; 3 % (3/97) and 1 % (6/422) for Babesia spp.; and 9 % (9/97) and 5 % (20/422) for Bartonella spp. Overall, between the five cities, there was no significant difference in the prevalence of any of the pathogens for the respective ticks (p > 0.05). The prevalence of B. burgdorferi s. l. in the questing and engorged I. ricinus varied from 0 to 27 % and 14-44%, respectively, with no statistical significance identified between the five cities (p > 0.05). In addition to reporting the updated data for Kyiv and Ternopil, this study is the first to provide the prevalences of the tick-borne pathogens for Chernivtsi, Khmelnytskyi, and Vinnytsia. This investigation is also the first to detect Neoehrlichia mikurensis in ixodid ticks from Ukraine. These new data will be useful for medical and veterinary practitioners as well as public health officials when diagnosing infections and when implementing measures to combat tick-borne diseases in Ukraine.
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Infecções por Anaplasmataceae/veterinária , Babesiose/epidemiologia , Infecções por Bartonella/veterinária , Doença de Lyme/veterinária , Infecções por Rickettsia/veterinária , Doenças Transmitidas por Carrapatos/veterinária , Anaplasmataceae/isolamento & purificação , Infecções por Anaplasmataceae/epidemiologia , Infecções por Anaplasmataceae/microbiologia , Animais , Animais Domésticos , Animais Selvagens , Babesia/isolamento & purificação , Babesiose/parasitologia , Bartonella/isolamento & purificação , Infecções por Bartonella/epidemiologia , Infecções por Bartonella/microbiologia , Grupo Borrelia Burgdorferi/isolamento & purificação , Dermacentor/microbiologia , Dermacentor/parasitologia , Ixodes/microbiologia , Ixodes/parasitologia , Doença de Lyme/epidemiologia , Doença de Lyme/microbiologia , Plantas , Prevalência , Rickettsia/isolamento & purificação , Infecções por Rickettsia/epidemiologia , Infecções por Rickettsia/microbiologia , Doenças Transmitidas por Carrapatos/epidemiologia , Doenças Transmitidas por Carrapatos/microbiologia , Doenças Transmitidas por Carrapatos/parasitologia , Ucrânia/epidemiologiaRESUMO
In Slovakia, little knowledge is available on the occurrence, hosts and vectors of Borrelia miyamotoi of the relapsing fever group. In the current study, 2160 questing and rodent-attached ticks of six species (Ixodes ricinus, Ixodes trianguliceps, Dermacentor marginatus, Dermacentor reticulatus, Haemaphysalis concinna and Haemaphysalis inermis), 279 fleas belonging to 9 species (Ctenophthalmus agyrtes, Ctenophthalmus solutus, Ctenophthalmus assimilis, Megabothris turbidus, Amalareus penicilliger, Hystrichopsylla orientalis, Ctenophthalmus uncinatus, Doratopsylla dasycnema and Nosopsyllus fasciatus) and skin biopsies from 245 small mammals belonging to eight species (Apodemus agrarius, Apodemus flavicollis, Apodemus uralensis, Myodes glareolus, Crocidura leucodon, Micromys minutus, Microtus arvalis, Microtus subterraneus) were screened for the presence of B. miyamotoi DNA. The overall prevalence of B. miyamotoi found in questing and rodent-attached ticks was 1.8% (23 positive/1260 examined) and 3.4% (31 positive/900 examined), respectively. Borrelia miyamotoi was detected in questing I. ricinus, rodent-attached I. ricinus and H. inermis ticks, and in one male of the common vole (M. arvalis) in different habitats (mainly rural) in eastern Slovakia. However, B. miyamotoi was not found in any of the tested fleas. Our findings indicate that rural habitats with different species of tick vectors and hosts are appropriate for the occurrence of B. miyamotoi.
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Infecções por Borrelia/veterinária , Borrelia/isolamento & purificação , Interações Hospedeiro-Parasita , Ixodidae/microbiologia , Doenças dos Roedores/epidemiologia , Sifonápteros/microbiologia , Animais , Infecções por Borrelia/epidemiologia , Infecções por Borrelia/microbiologia , Meio Ambiente , Prevalência , Doenças dos Roedores/microbiologia , Roedores , EslováquiaRESUMO
Lyme borreliosis and tick-borne encephalitis (TBE) are the most common tick-borne diseases in Germany. We collected Ixodes ricinus ticks from 16 high-risk and four low-risk sites distributed in Lower Bavaria and Upper Palatinate based on the number of human TBE cases recorded at the Robert Koch Institute from 2001 to 2009. A total of 8805 questing ticks (8203 nymphs, 602 adults) were collected in 2010 and examined in pools for the presence of tick-borne encephalitis virus (TBEV) using real-time RT-PCR. Overall TBEV prevalence evaluated as the minimum infection rate (MIR) was 0.26 % (23 positive pools/8805 ticks in 1029 pools). TBEV was detected at seven of the 16 high-risk sites, where MIR ranged from 0.16 to 2.86 %. A total of 3969 ticks were examined by PCR for infection with Borrelia burgdorferi sensu lato (s.l.) targeting the 5â¯S-23â¯S rRNA intergenic spacer (IGS) region. IGS nucleotide sequences were used to determine genospecies. Selected positive Borrelia samples were subjected to PCR and sequencing targeting the OspA gene, providing 46 sequences for molecular phylogenetic analysis. Of the 3969 questing ticks, 506 (12.7 %) were positive for B. burgdorferi s.l. Seven B. burgdorferi s.l. genospecies were identified: B. afzelii (41.3 %), B. garinii (19 %), B. valaisiana (13.8 %), B. burgdorferi sensu stricto (11.1 %), B. spielmanii (0.4 %), B. lusitaniae (0.2 %), and Candidatus B. finlandensis (0.6 %). Mixed infections were identified in 13.6 % of the ticks. The rate of infection in questing ticks varied among sites from 5.6 % (72 examined, four positive) to 29.5 % (88 examined, 26 positive). B. burgdorferi s.l. occurred at all 20 sites, whereas TBEV was detected only at the high-risk sites where more human TBE cases were reported compared to low-risk sites.
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Grupo Borrelia Burgdorferi/isolamento & purificação , Vírus da Encefalite Transmitidos por Carrapatos/isolamento & purificação , Ixodes/microbiologia , Animais , Feminino , Alemanha , Ixodes/virologia , Masculino , Ninfa , Prevalência , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Research projects in the field of eco-epidemiology of tick-borne diseases often require extensive sampling of arthropod vectors in the field. The aim of our study was to use geographical information systems (GIS) to select appropriate sampling sites of Ixodes ricinus ticks in central European habitat for further ecological studies of vector-borne pathogens (tick-borne encephalitis virus and Borrelia burgdorferi sensu lato). The model area was the Czech-German borderland (the region of South Bohemia and two regions in Germany: the Upper Palatinate and Lower Bavaria) where numerous human tick-borne encephalitis cases are reported annually. We prepared the sampling site design as a multi-criteria evaluation (MCE) task. In the GIS environment, we conducted MCE with a set of environmental, socio-economic and epidemiological data (altitude, vegetation cover, number of tick-borne encephalitis cases recorded in the past, tourist activity). The MCE classified the surveyed area into two classes: suitable for tick collection and unsuitable for tick collection. Subsequently, 50 tick sampling sites were randomly selected in the suitable area: 30 in South Bohemia (Czech Republic) and 20 in the Upper Palatinate and Lower Bavaria regions (Bavaria, Germany). The sampling sites were identified and surveyed in the field. The presence of ticks was confirmed by flagging at each of the selected plots. The described MCE system represents a versatile tool for semi-randomized design of tick sampling sites for research projects in the field of tick-borne pathogen ecology as well as for tick-borne pathogen surveillance programs run by local health authorities.
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Ecossistema , Ixodes/fisiologia , Doenças Transmitidas por Carrapatos/epidemiologia , Altitude , Animais , Borrelia burgdorferi/isolamento & purificação , Vírus da Encefalite Transmitidos por Carrapatos/isolamento & purificação , Feminino , Sistemas de Informação Geográfica , Geografia , Masculino , Estudos de AmostragemRESUMO
Monitoring West Nile virus (WNV) and Usutu virus (USUV) activity now has the highest priority among mosquito-borne pathogenic viruses circulating in the European Union. This study documents a first time detection and the co-circulation of WNV lineage-2 (with the minimal prevalence of 0.46%) and USUV clade Europe 2 (with the minimal prevalence of 0.25%) in mosquitoes from the same habitat of south-western Slovakia and underlines necessity to perform rigorous surveillance in birds, mosquitoes, horses and humans in that country.
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Culicidae/virologia , Infecções por Flavivirus/veterinária , Flavivirus/isolamento & purificação , Febre do Nilo Ocidental/veterinária , Vírus do Nilo Ocidental/isolamento & purificação , Animais , Aves/virologia , Culex/virologia , Ecossistema , Feminino , Flavivirus/genética , Infecções por Flavivirus/epidemiologia , Cavalos/virologia , Humanos , Mosquitos Vetores/virologia , Filogenia , Prevalência , Saúde Pública , Eslováquia/epidemiologia , Febre do Nilo Ocidental/epidemiologia , Vírus do Nilo Ocidental/genéticaRESUMO
As a part of a larger survey a total of 599 Ixodes ricinus ticks collected from four locations (Neustadt an der Waldnaab, Amberg, Poppenricht and Lintach) in the north of the Upper Palatinate in Bavaria were investigated for infection with Borrelia burgdorferi sensu lato (s. l.) species using the 5S-23S intergenic spacer of rRNA gene as a target in a nested PCR (Rijpkema et al., 1995) and a sequencing method. Overall, 15.8% ticks were infected with B. burgdorferi s. I. Borrelia afzelii (43.1%) was the predominant genospecies, followed by Borrelia valaisiana (14.7%), Borrelia garinii (13.7%) and Borrelia burgdorferi sensu stricto (6.3%). Also Borrelia spielmanii was found (1.1%). Of the infected ticks, 21.1% harbored multiple infections with B. burgdorferi s. I. The highest number of infected ticks was found in Amberg (22.5%) and the lowest number in Neustadt an der Waldnaab (11.9%). In Poppenricht and Lintach, the numbers of infected ticks were 12% and 18.7%, respectively. Human pathogenic Borrelia species were found to be prevalent at each study site thus representing the potential risk for people living and visiting these areas.