Ticks are important vectors of various microorganisms, including bacteria. In this study, we examined Hyalomma aegyptium ticks collected from 240 spur-thighed tortoises Testudo graeca at 42 localities in the Mediterranean and Middle East and analysed them for the presence of bacteria of the genera Anaplasma, Borrelia, Coxiella, and Rickettsia. Altogether, 576 out of 928 analysed ticks (62.1%) were positive for at least one of the tested bacteria. The highest prevalence in individual ticks was found for Borrelia turcica (43.6%), followed by Rickettsia (12.3%) and Anaplasma (6.1%). No sample was positive for Coxiella burnetii. Among Rickettsia, we detected two species, Rickettsia africae and Rickettsia aeschlimannii, and also other unspecified Rickettsia. Anaplasma (100% identity with A. phagocytophilum) was detected at 15 (35%) out of 42 studied localities, any of Rickettsia at 28 (67%), and B. turcica at 32 (76%) localities. The geographic distribution of the studied microorganisms varied, with none of them detected in Syria, and only Rickettsia spp. detected in Morocco. Sequence analysis revealed substantial genetic variability in all detected agents, with the most variable (36 new haplotypes) being glpQ gene used as a marker for B. turcica. We also analysed the prevalence of various co-infections among studied ticks, with the mean number of co-infected ticks per tortoise increased with the number of ticks per tortoise. However, the frequencies of co-infected ticks do not indicate the presence of antagonistic or synergistic facilitative interactions between the agents. According to our data, we could expect that the eco-epidemiological importance of H. aegyptium does not stem from their tortoise hosts but rather from the low host specificity of its larvae and nymphs, feeding on a wider spectrum of reptilian, avian, and mammalian hosts.

• MeSH
• Anaplasma * isolation & purification   MeSH
• Borrelia isolation & purification   MeSH
• Coxiella isolation & purification   genetics   MeSH
• Tick Infestations veterinary   epidemiology   parasitology   MeSH
• Ixodidae * microbiology   growth & development   MeSH
• Rickettsia * isolation & purification   MeSH
• Turtles * microbiology   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Middle East MeSH
• Mediterranean Region MeSH

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 isolation & purification   genetics   MeSH
• Phylogeny * MeSH
• Tick Infestations * veterinary   parasitology   MeSH
• Cats parasitology   MeSH
• Tick-Borne Diseases veterinary   microbiology   parasitology   MeSH
• Dog Diseases * parasitology   diagnosis   MeSH
• Dogs MeSH
• Rhipicephalus sanguineus * microbiology   MeSH
• Rickettsia conorii isolation & purification   genetics   MeSH
• RNA, Ribosomal, 16S analysis   genetics   MeSH
• RNA, Ribosomal * MeSH
• Animals MeSH
• Check Tag
• Cats parasitology   MeSH
• Male MeSH
• Dogs MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Case Reports MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Hungary 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
• Ixodes * MeSH
• Mosquito Vectors MeSH
• Rickettsia * MeSH
• Mammals MeSH
• Seroepidemiologic Studies MeSH
• Encephalitis Viruses, Tick-Borne * MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Europe 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
• Cell Line MeSH
• Arthropods * MeSH
• Ixodes * microbiology   MeSH
• Rickettsia felis * genetics   MeSH
• Rickettsia * genetics   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't 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 isolation & purification   MeSH
• Babesia isolation & purification   MeSH
• Ixodidae microbiology   parasitology   MeSH
• Rickettsia isolation & purification   MeSH
• Environment MeSH
• Zoonoses * microbiology   parasitology   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Czech Republic 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
• Rocky Mountain Spotted Fever epidemiology   MeSH
• Ticks microbiology   MeSH
• Cats MeSH
• Humans MeSH
• Cat Diseases * diagnosis   epidemiology   microbiology   MeSH
• Dog Diseases * diagnosis   epidemiology   microbiology   MeSH
• Dogs MeSH
• Rhipicephalus sanguineus * MeSH
• Rickettsia MeSH
• Rickettsia Infections * diagnosis   epidemiology   veterinary   MeSH
• Seroepidemiologic Studies MeSH
• Animals MeSH
• Check Tag
• Cats MeSH
• Humans MeSH
• Dogs MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Brazil MeSH

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.; 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.

• MeSH
• Anaplasma pathogenicity   MeSH
• Babesia pathogenicity   MeSH
• Humans MeSH
• Mobile Applications MeSH
• Multiplex Polymerase Chain Reaction MeSH
• Tick-Borne Diseases epidemiology   prevention & control   MeSH
• Disease Hotspot MeSH
• Primary Prevention MeSH
• Rickettsia pathogenicity   MeSH
• Risk Factors MeSH
• Check Tag
• Humans MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• epidemiologie
• infekční lékařství
• NML Publication type
• 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
• Chromatography, Liquid MeSH
• Rabbits MeSH
• Humans MeSH
• Models, Molecular MeSH
• Molecular Weight MeSH
• Bacterial Outer Membrane Proteins chemistry   immunology   metabolism   MeSH
• Proteomics methods   MeSH
• Antibodies, Bacterial blood   MeSH
• Rickettsia akari immunology   isolation & purification   metabolism   MeSH
• Protein Structure, Secondary MeSH
• Spotted Fever Group Rickettsiosis diagnosis   immunology   MeSH
• Tandem Mass Spectrometry MeSH
• Animals MeSH
• Check Tag
• Rabbits MeSH
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't 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 isolation & purification   MeSH
• DNA, Bacterial analysis   MeSH
• Hydrolysis MeSH
• Ixodes microbiology   MeSH
• Polymerase Chain Reaction methods   MeSH
• Rickettsia isolation & purification   MeSH
• Sensitivity and Specificity MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't 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
• Ticks microbiology   MeSH
• Dog Diseases diagnosis   microbiology   MeSH
• Polymerase Chain Reaction MeSH
• Antibodies, Bacterial blood   MeSH
• Dogs MeSH
• Rickettsia classification   genetics   isolation & purification   MeSH
• Spotted Fever Group Rickettsiosis diagnosis   microbiology   veterinary   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Dogs MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Brazil MeSH