The brown dog tick, Rhipicephalus sanguineus is a complex of tick species with an unsettled species concept. In Europe, R. sanguineus is considered mainly a Mediterranean tick with sporadic findings in central and northern Europe. R. sanguineus is known as a vector of a range of pathogens of medical and veterinary importance, most of which not yet reported as autochthonous in Hungary. A total of 1839 ticks collected by veterinarians from dogs and cats were obtained in Hungary. The study aims at precise determination of ticks identified as R. sanguineus and detection of pathogens in collected ticks. All ticks were morphologically determined and 169 individuals were identified as R. sanguineus. A subset of 15 ticks was selected for molecular analysis (16S rDNA, 12S rDNA, COI). Phylogenetic analyses invariably placed sequences of all three markers into a single haplotype identified as R. sanguineus sensu stricto. All 169 brown dog ticks were tested for the presence of A. platys, E. canis, R. conorii, B. vogeli and H. canis. None of the investigated ticks was positive for the screened pathogens, though A. phagocytophilum sequence was detected in a single tick.

• MeSH
• Anaplasma * MeSH
• Ehrlichia canis izolace a purifikace   genetika   MeSH
• fylogeneze * MeSH
• infestace klíšťaty * veterinární   parazitologie   MeSH
• kočky parazitologie   MeSH
• nemoci přenášené klíšťaty veterinární   mikrobiologie   parazitologie   MeSH
• nemoci psů * parazitologie   diagnóza   MeSH
• psi MeSH
• Rhipicephalus sanguineus * mikrobiologie   MeSH
• Rickettsia conorii izolace a purifikace   genetika   MeSH
• RNA ribozomální 16S analýza   genetika   MeSH
• RNA ribozomální * MeSH
• zvířata MeSH
• Check Tag
• kočky parazitologie   MeSH
• mužské pohlaví MeSH
• psi MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Maďarsko MeSH

BACKGROUND: Vector-borne zoonotic diseases are a concerning issue in Europe. Lyme disease and tick-borne encephalitis virus (TBEV) have been reported in several countries with a large impact on public health; other emerging pathogens, such as Rickettsiales, and mosquito-borne flaviviruses have been increasingly reported. All these pathogens are linked to wild ungulates playing roles as tick feeders, spreaders, and sentinels for pathogen circulation. This study evaluated the prevalence of TBEV, Borrelia burgdorferi sensu lato, Rickettsia spp., Ehrlichia spp., and Coxiella spp. by biomolecular screening of blood samples and ticks collected from wild ungulates. Ungulates were also screened by ELISA and virus neutralization tests for flaviviral antibody detection. RESULTS: A total of 274 blood samples were collected from several wild ungulate species, as well as 406 Ixodes ricinus, which were feeding on them. Blood samples tested positive for B. burgdorferi s.l. (1.1%; 0-2.3%) and Rickettsia spp. (1.1%; 0-2.3%) and showed an overall flaviviral seroprevalence of 30.6% (22.1-39.2%): 26.1% (17.9-34.3%) for TBEV, 3.6% (0.1-7.1%) for Usutu virus and 0.9% (0-2.7%) for West Nile virus. Ticks were pooled when possible and yielded 331 tick samples that tested positive for B. burgdorferi s.l. (8.8%; 5.8-11.8%), Rickettsia spp. (26.6%; 21.8-31.2%) and Neoehrlichia mikurensis (1.2%; 0-2.4%). TBEV and Coxiella spp. were not detected in either blood or tick samples. CONCLUSIONS: This research highlighted a high prevalence of several tick-borne zoonotic pathogens and high seroprevalence for flaviviruses in both hilly and alpine areas. For the first time, an alpine chamois tested positive for anti-TBEV antibodies. Ungulate species are of particular interest due to their sentinel role in flavivirus circulation and their indirect role in tick-borne diseases and maintenance as Ixodes feeders and spreaders.

• MeSH
• Coxiella MeSH
• klíště * MeSH
• komáří přenašeči MeSH
• Rickettsia * MeSH
• savci MeSH
• séroepidemiologické studie MeSH
• viry klíšťové encefalitidy * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa MeSH

Although the cat flea, Ctenocephalides felis, has been identified as the primary vector of Rickettsia felis, additional flea, tick, mite, and louse species have also been associated with this bacterium by molecular means; however, the role of these arthropods in the transmission of R. felis has not been clarified. Here, we succeeded in culture isolation of R. felis from a host-seeking castor bean tick, Ixodes ricinus, the most common tick in Slovakia. The bacterial isolation was performed on XTC-2 cells at 28 °C using the shell-vial technique. An evaluation of the growth properties was performed for both the XTC-2 and Vero cell lines. We observed R. felis in the infected host cells microscopically by Gimenez staining and immunofluorescence assay. The R. felis isolate was purified by gradient ultracentrifugation and visualized by electron microscopy. Fragments of the genes gltA, ompA, ompB, htrA, rpoB, sca4, rffE, and rrs were amplified and compared with the corresponding sequences of the type strain URRWXCal2 and other R. felis culture -isolated strains. We did not detect any nucleotide polymorphisms; however, plasmid pRFδ, characteristic of the standard strain, was absent in our isolate. Herein, we describe the first successful isolation and characterization of a tick-derived R. felis strain "Danube", obtained from an I. ricinus nymph.

• MeSH
• buněčné linie MeSH
• členovci * MeSH
• klíště * mikrobiologie   MeSH
• Rickettsia felis * genetika   MeSH
• Rickettsia * genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Studies of tick-borne diseases (TBDs) in Europe focus on pathogens with principal medical importance (e.g. Lyme disease and tick-borne encephalitis), but we have limited epidemiological information on the neglected pathogens, such as the members of the genera Anaplasma, Rickettsia, Babesia and Candidatus Neoehrlichia mikurensis. Here, we integrated an extensive field sampling, laboratory analysis and GIS models to provide first publicly available information on pathogen diversity, prevalence and infection risk for four overlooked zoonotic TBDs in the Czech Republic. In addition, we assessed the effect of landscape variables on the abundance of questing ticks at different spatial scales and examined whether pathogen prevalence increased with tick density. Our data from 13,340 ticks collected in 142 municipalities showed that A. phagocytophilum (MIR = 3.5%) and Ca. Neoehrlichia mikurensis (MIR = 4.0%) pose geographically uneven risks with localized hotspots, while Rickettsia (MIR = 4.9%) and Babesia (MIR = 1.1%) had relatively homogeneous spatial distribution. Landscape variables had significant effect on tick abundance up to the scale of 1 km around the sampling sites. Questing ticks responded positively to landscape diversity and configuration, especially to forest patch density that strongly correlates with the amount of woodland-grassland ecotones. For all four pathogens, we found higher prevalence in places with higher densities of ticks, confirming the hypothesis that tick abundance amplifies the risk of TB infection. Our findings highlight the importance of landscape parameters for tick vectors, likely due to their effect on small vertebrates as reservoir hosts. Future studies should explicitly investigate the combined effect of landscape parameters and the composition and population dynamics of hosts on the host-vector-pathogen system.

• MeSH
• Anaplasmataceae izolace a purifikace   MeSH
• Babesia izolace a purifikace   MeSH
• Ixodidae mikrobiologie   parazitologie   MeSH
• Rickettsia izolace a purifikace   MeSH
• životní prostředí MeSH
• zoonózy * mikrobiologie   parazitologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

Spotted fever group rickettsioses are emerging diseases. In some of these diseases, domestic dogs act as sentinels. Canine serological studies have demonstrated that rickettsial dispersion is concentrated in rural areas, seroprevalence being higher where human rickettsioses are endemic. In Rio de Janeiro, the Atlantic forest vegetation has been devastated by urbanization. In this context, we aimed to detect Rickettsia spp. in urban areas of the West Zone of Rio de Janeiro. Sera from 130 dogs were tested by Indirect Immunofluorescence Assay, and ticks collected from these dogs were tested by polymerase chain reaction. We found the rate of serological reactions against R. rickettsii and R. parkeri in our study area to exceed those of rural and non-endemic areas, highlighting the importance of dogs as urban sentinels. The possibility of contact with opossums and capybaras increased the chances of exposure to Rickettsia spp., reinforcing the hypothetical link between the landscape and the rickettsial wild cycle. Rhipicephalus sanguineus sensu lato was the tick most frequently observed. PCR-positive samples showed similarity with R. rickettsii and R. felis, an emerging pathogen rarely reported from ticks. We observed that rickettsiae circulate in urban places and ticks from indoor environments, which may be involved in bacterial epidemiology.

• MeSH
• horečka Skalistých hor epidemiologie   MeSH
• klíšťata mikrobiologie   MeSH
• kočky MeSH
• lidé MeSH
• nemoci koček * diagnóza   epidemiologie   mikrobiologie   MeSH
• nemoci psů * diagnóza   epidemiologie   mikrobiologie   MeSH
• psi MeSH
• Rhipicephalus sanguineus * MeSH
• Rickettsia MeSH
• rickettsiové infekce * diagnóza   epidemiologie   veterinární   MeSH
• séroepidemiologické studie MeSH
• zvířata MeSH
• Check Tag
• kočky MeSH
• lidé MeSH
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Brazílie MeSH

The project aims to investigate distribution, abundance, and prevalence of pathogenic agents of the genera Rickettsia, Anaplasma and Babesia and using these models focus two main objectives. First of them is to prepare, applying GIS modelling, epidemiological risk maps for these agents in the Czech Republic. Preparation of publicly available platform useful also in future analyses of distribution and abundance of various agents represents the second objective. To achieve this, the project employs complex approach, including study of evolutionary relationships of particular collected isolates and their taxonomy. Establishment of fast, specific, sensitive, and reliable quantitative diagnostic method is essential part of the project. Biodiversity of studied model groups in the Czech Republic will be also clarified. Obtained results will be available as platform for following studies of tick-borne pathogenic agents, including yet neglected groups. Complementary skills of both cooperating teams are indisputably useful advantage of the proposed project.

Projekt sleduje pomocí studia distribuce, abundance a prevalence třech skupin relativně přehlížených, klíšťaty přenosných patogenů, (a sice zástupců rodů Rickettsia, Anaplasma a Babesia) dva hlavní cíle. Prvním z nich je prostřednictvím modelování v prostředí GIS vytvořit české mapy rizik pro výše zmíněné skupiny patogenních agens. Druhým cílem je tvorba veřejně přístupné aplikace pro možnost následných analýz rozšíření a početnosti různých studovaných patogenů. K dosažení těchto cílů projekt využívá komplexní přístup, včetně studia evolučních vztahů jednotlivých získaných izolátů a jejich taxonomie. Nezbytnou součástí projektu je příprava rychlé, specifické, citlivé a spolehlivé kvantitativní diagnostické metodiky. V projektu bude objasněna také biodiverzita studovaných skupin ve vztahu k ČR. Získané výsledky poslouží jako platforma pro další studium klíšťaty přenosných patogenů, včetně dosud opomíjených skupin. Metodicky projekt využívá výhod komplementárních dovedností dvou spolupracujících týmů.

• MeSH
• Anaplasma patogenita   MeSH
• Babesia patogenita   MeSH
• lidé MeSH
• mobilní aplikace MeSH
• multiplexová polymerázová řetězová reakce MeSH
• nemoci přenášené klíšťaty epidemiologie   prevence a kontrola   MeSH
• ohnisko nemoci MeSH
• primární prevence MeSH
• Rickettsia patogenita   MeSH
• rizikové faktory MeSH
• Check Tag
• lidé MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• epidemiologie
• infekční lékařství
• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

BACKGROUND: Rickettsialpox is a febrile illness caused by the mite-borne pathogen Rickettsia akari. Several cases of this disease are reported worldwide annually. Nevertheless, the relationship between the immunogenicity of R. akari and disease development is still poorly understood. Thus, misdiagnosis is frequent. Our study is aiming to identify immunogenic proteins that may improve disease recognition and enhance subsequent treatment. To achieve this goal, two proteomics methodologies were applied, followed by immunoblot confirmation. RESULTS: Three hundred and sixteen unique proteins were identified in the whole-cell extract of R. akari. The most represented protein groups were found to be those involved in translation, post-translational modifications, energy production, and cell wall development. A significant number of proteins belonged to amino acid transport and intracellular trafficking. Also, some proteins affecting the virulence were detected. In silico analysis of membrane enriched proteins revealed 25 putative outer membrane proteins containing beta-barrel structure and 11 proteins having a secretion signal peptide sequence. Using rabbit and human sera, various immunoreactive proteins were identified from which the 44 kDa uncharacterized protein (A8GP63) has demonstrated a unique detection capability. It positively distinguished the sera of patients with Rickettsialpox from other rickettsiae positive human sera. CONCLUSION: Our proteomic analysis certainly contributed to the lack of knowledge of R. akari pathogenesis. The result obtained may also serve as a guideline for a more accurate diagnosis of rickettsial diseases. The identified 44 kDa uncharacterized protein can be certainly used as a unique marker of rickettsialpox or as a target molecule for the development of more effective treatment.

• MeSH
• chromatografie kapalinová MeSH
• králíci MeSH
• lidé MeSH
• molekulární modely MeSH
• molekulová hmotnost MeSH
• proteiny vnější bakteriální membrány chemie   imunologie   metabolismus   MeSH
• proteomika metody   MeSH
• protilátky bakteriální krev   MeSH
• Rickettsia akari imunologie   izolace a purifikace   metabolismus   MeSH
• sekundární struktura proteinů MeSH
• skvrnité horečky diagnóza   imunologie   MeSH
• tandemová hmotnostní spektrometrie MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Anaplasma phagocytophilum and Rickettsia spp. are vector-borne zoonotic bacteria, which are clinically important especially in immunocompromised patients. There are large gaps in the current knowledge of their geographic distribution and prevalence in both their vectors and hosts. Our aim was to develop reliable and easy detection method for both these pathogens. We made a new hydrolysis probe based duplex Real-Time PCR assay based on previous studies. We optimized the assays and tested them to provide reliable recommended procedures with a sensitivity to a minimum of 10 target DNA copies per sample. The assays were designed to be specific for A. phagocytophilum and in the same reaction detect multiple species of rickettsiae. We designed gBlock quantification standards that provide the option to identify differences in pathogen load among different samples in subsequent studies.

• MeSH
• Anaplasma phagocytophilum izolace a purifikace   MeSH
• DNA bakterií analýza   MeSH
• hydrolýza MeSH
• klíště mikrobiologie   MeSH
• polymerázová řetězová reakce metody   MeSH
• Rickettsia izolace a purifikace   MeSH
• senzitivita a specificita MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Rickettsia rickettsii is the causative agent of Brazilian spotted fever (BSF), for which humans and dogs are both susceptible. Dogs are sentinels in serological surveys, however, canine disease is rarely reported. Therefore, we aimed to evaluate natural infection by spotted fever group (SFG) Rickettsia spp. in dogs and ticks collected from domiciles close to forest fragments, featuring domestic-wildlife interface areas. Samples from 115 dogs and 135 ixodids were assessed by polymerase chain reactions (PCR) targeting the gltA gene for Rickettsia spp. and the ompA gene for the SFG rickettsial species. One dog (0.87%; 1/115) was positive for R. rickettsii. This dog presented nonspecific laboratory and clinical abnormalities (thrombocytopenia, hyperproteinemia, lymph node enlargement, emaciation, anorexia, and lethargy). Rickettsia parkeri was identified in 2.96% (4/135) of the ticks (Amblyomma sculptum, A. aureolatum, and Rhipicephalus sanguineus). This study confirmed the presence of SFG bacteria in non-endemic and preserved locations, where domestic and wild populations interact. We reinforce the fact that the dog is susceptible to natural R. rickettsii infection. Although this is a rare finding, preventive measures should be taken against BSF in the studied areas. Finally, R. parkeri infection is possibly being demonstrated in A. sculptum for the first time.

• MeSH
• klíšťata mikrobiologie   MeSH
• nemoci psů diagnóza   mikrobiologie   MeSH
• polymerázová řetězová reakce MeSH
• protilátky bakteriální krev   MeSH
• psi MeSH
• Rickettsia klasifikace   genetika   izolace a purifikace   MeSH
• skvrnité horečky diagnóza   mikrobiologie   veterinární   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Brazílie MeSH

Companion animals can be infested by various species of parasitic insects. Cat flea Ctenocephalides felis (C. felis felis) (Bouché, 1835) and dog flea Ctenocephalides canis (Curtis, 1826) belong to multihost external parasites of mammals, which most frequently occur on domestic cats Felis catus Linnaeus and dogs Canis familiaris Linnaeus. The main aim of this study was to investigate the presence of pathogens, such as Anaplasma phagocytophilum (syn. Ehrlichia phagocytophila) and Rickettsia spp., in adult C. felis and C. canis fleas. Flea sampling has been realised from January 2013 to April 2017 in veterinary clinics, animal shelters and pet grooming salons. Fleas were collected from domestic cats and dogs, directly from the pet skin or hair. Then, the DNA was isolated from a single flea by using the alkaline hydrolysis and samples were screened for the presence of pathogens using PCR method. Anaplasma phagocytophilum has occurred in 29% of examined C. felis and 16% of C. canis individuals. In turn, the prevalence of Rickettsia spp. in cat fleas population was only 3%, and the dog fleas 7%. The present study showed the presence of pathogenic agents in cat and dog fleas, which indicates the potential role of these insects in circulation of A. phagocytophilum and Rickettsia spp. in the natural habitat. Furthermore, exposition to these flea species, whose hosts are domestic cats and dogs, can pose a potential risk of infection for humans.

• MeSH
• Anaplasma phagocytophilum izolace a purifikace   MeSH
• Ctenocephalides mikrobiologie   MeSH
• Rickettsia izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Polsko MeSH