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

The Gram-negative, obligate intracellular tick-transmitted pathogen Anaplasma phagocytophilum can cause acute febrile diseases in humans and domestic animals. The expansion of the tick Ixodes ricinus (Linnaeus, 1758) in northern Europe due to climate change is of serious concern for animal and human health. The aim of the present study was to investigate the impact of A. phagocytophilum infection in moose Alces alces (Linnaeus) calves by evaluating the carcass weights of infected and non-infected animals and examining animal tissues samples for co-infections with either species of Babesia Starcovici, 1893 or bacteria of the genus Bartonella. The carcasses of 68 free-ranging moose calves were weighed by hunters during the hunting seasons from 2014 to 2017 in two regions in southern Norway and spleen samples were collected. Anaplasma phagocytophilum was detected in moose sampled from locations infected with ticks with a prevalence of 82% (n = 46). The carcass weights of A. phagocytophilum-infected calves (n = 46) and non-infected (n = 22) calves were compared. Although the average weight of infected calves (45.6 kg) was lower than that of non-infected calves (46.5 kg), the difference was not statistically significant. Three different variants of the bacterium 16S rRNA gene were identified. The average weight of animals infected with variant I was 49.9 kg, whereas that of animals infected with variant III was 42.0 kg, but the difference was not statistically significant (p = 0.077). Co-infections of A. phagocytophilum with Bartonella spp. or with Babesia spp. were found in 20 and two calves, respectively. A triple infection was found in two calves. Sequence analysis of the 18S rRNA gene of Babesia-positive samples revealed the presence of Babesia cf. odocoilei (Emerson et Wright, 1970). Strains of Bartonella closely related to Bartonella bovis (Bermond, Boulouis, Heller, Laere, Monteil, Chomel, Sander, Dehio et Piemont, 2002) were identified based on phylogenetic analysis of the gltA and rpoB genes. The loss of body mass in moose calves in the tick-infected site was probably influenced by multiple factors.

• MeSH
• Anaplasma phagocytophilum * klasifikace   genetika   izolace a purifikace   MeSH
• Babesia genetika   MeSH
• Bartonella genetika   MeSH
• DNA bakterií chemie   genetika   izolace a purifikace   MeSH
• ehrlichióza komplikace   epidemiologie   patologie   veterinární   MeSH
• fylogeneze MeSH
• oligonukleotidy chemie   MeSH
• polymerázová řetězová reakce veterinární   MeSH
• sekvence nukleotidů MeSH
• slezina mikrobiologie   patologie   MeSH
• tělesná hmotnost MeSH
• vysoká zvěř * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Norsko 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

The wild boar (Sus scrofa) population has increased dramatically over the last decades throughout Europe and it has become a serious pest. In addition, the common habitat of wild boar and of the tick, Ixodes ricinus, indicates the potential of wild boar to play a role in epidemiology of epizootic and zoonotic tick-borne pathogens, including Anaplasma phagocytophilum. In Europe, epidemiological cycles and reservoirs of A. phagocytophilum, including its zoonotic haplotypes, are poorly understood. In this study, we focused on detection and further genetic characterization of A. phagocytophilum and piroplasmids in 550 wild boars from eleven districts of Moravia and Silesia in the Czech Republic. Using highly sensitive nested PCR targeting the groEL gene, the DNA of A. phagocytophilum was detected in 28 wild boars (5.1 %) representing six unique haplotypes. The dominant haplotype was found in 21 samples from 7 different districts. All detected haplotypes clustered in the largest clade representing the European ecotype I and the dominant haplotype fell to the subclade with the European human cases and strains from dogs and horses. Nested PCR targeting the variable region of the 18S rRNA gene of piroplasmids resulted in one positive sample with 99.8 % sequence identity to Babesia divergens. The presence of these two pathogens that are primarily circulated by I. ricinus confirms the local participation of wild boar in the host spectrum of this tick and warrants experimental studies to address wild boar as a reservoir of zoonotic haplotypes of A. phagocytophilum.

• MeSH
• Anaplasma phagocytophilum genetika   izolace a purifikace   MeSH
• anaplasmóza epidemiologie   mikrobiologie   MeSH
• babezióza epidemiologie   parazitologie   MeSH
• bakteriální geny MeSH
• genetická variace * MeSH
• nemoci prasat epidemiologie   mikrobiologie   parazitologie   MeSH
• Piroplasmida genetika   izolace a purifikace   MeSH
• prasata MeSH
• prevalence MeSH
• protozoální geny MeSH
• Sus scrofa MeSH
• zdroje nemoci parazitologie   veterinární   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

Interactions among endosymbiotic bacteria inside their eukaryotic hosts are poorly understood, particularly in mites. The mite Tyrophagus putrescentiae is a common, medically important generalist species that has many intracellular and gut bacterial symbionts. In the experiments, we examined bacterial abundances and composition in mite populations obtained by controlled mixing of stock mite populations that differed in the presence/absence of the major intracellular bacteria Wolbachia and Cardinium. Changes in microbial communities were characterized using 16S ribosomal RNA high-throughput sequencing (pooled mite individuals) and quantitative PCR for key microbial taxa (individual mites). Mite fitness was estimated as a parameter of population growth. We detected that in mixed mite populations, Cardinium and Wolbachia can co-occur in the same mite individual. The presence of Cardinium was negatively correlated with the presence of Wolbachia and Bartonella, while the Bartonella and Wolbachia were positively correlated in individual level samples. Since mixed populations had lower abundances of Wolbachia, while the abundance of Cardinium did not change, we suggest that the presence of Cardinium inhibits the growth of Wolbachia. The mixed mite populations had lower population growth than parental populations. The possible effect of symbionts on the fitness of mixed population is discussed.

• MeSH
• Acaridae * MeSH
• Bacteroidetes genetika   MeSH
• lidé MeSH
• mikrobiota * MeSH
• roztoči * MeSH
• symbióza MeSH
• Wolbachia * genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Tyrophagus putrescentiae is an astigmatid mite of great economic, medical and veterinary importance. The microbiome, especially intracellular bacteria, may affect allergy/allergen expression. We targeted Wolbachia proteins, allergen comparisons and markers in Wolbachia-mite interactions in three mite populations. A decoy database was constructed by proteogenomics using the T. putrescentiae draft genome, Wolbachia transcriptome assembly and current T. putrescentiae-related sequences in GenBank. Among thousands of mite-derived proteins, 18 Wolbachia proteins were reliably identified. We suggest that peroxiredoxin, bacterioferritin, ankyrin repeat domain-containing protein and DegQ family serine endoprotease indicate a higher-level bacterium-bacterium-host interaction. We produced evidence that the host-Wolbachia interaction is modulated through pattern recognition receptors (PRRs), mannose-binding lectins/mannose receptors, the cholinergic anti-inflammatory pathway with TNF-α, and others. We observed Tyr p 3 suppression in mites with Wolbachia, linking trypsin to PRR modulation. Nine out of the 12 current WHO/IUIS official allergens were reliably identified, but the remaining three allergens, Tyr p 1, 8 and 35, were detected as only trace hits. This study provides numerous markers for further Wolbachia-host interaction research. For accuracy, mite allergens should be considered according to abundance in species, but mite populations/strains, as well as their microbiome structure, may be key factors. SIGNIFICANCE: The astigmatid mites occurring in homes are significant producers of allergens that are highly dangerous to humans and domesticated animals. Mites are tightly associated with microorganisms that affect their biology and consequently allergy signatures. Mite populations were found to be infected with certain intracellular bacteria, but some populations lacked an intracellular bacterium. Our previous research showed that some populations of Tyrophagus putrescentiae are infected with Wolbachia, but some populations host additional bacteria of interest. Thus, there are not only interactions between the mites and Wolbachia but also likely an additional level of interaction that can be found in the interaction between different bacteria in the mites. These "higher-level" signatures and consequences that bacteria affect, including allergen production, are not understood in mites. In this study, we identified Wolbachia-specific proteins in mites for the first time. This study provides Wolbachia- and mite-derived markers that can be clues for describing "higher-level" mite-bacterium-bacterium interactions. Indeed, the microbiome contribution to allergies can potentially be derived directly from bacterial proteins, especially if they are abundant.

• MeSH
• alergeny * MeSH
• proteom MeSH
• proteomika MeSH
• roztoči * mikrobiologie   MeSH
• Wolbachia * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

European brown hare (Lepus europaeus Pallas 1778) is a broadly distributed lagomorph species in Europe, recognized as a host for Ixodes ricinus and reservoir of a wide range of pathogens with zoonotic potential. Even though Lepus europaeus represents an important game animal in Central Europe, the data available on Anaplasma phagocytophilum in this lagomorph are scarce. In this study, three populations of brown hare from distinct localities in the Czech Republic were analysed for the presence of Anaplasma phagocytophilum DNA. We used standard qPCR, targeting the msp2 gene and adapted the same assay also for digital droplet PCR. Out of 91 samples, these two methods identified 9 and 12 as positive, respectively. For taxonomic analysis, we amplified the groEL gene from five of six samples that were found positive by both methods. In phylogenetic analyses, this haplotype belongs to ecotype 1, and to the subclade with isolates from cervids and I. ricinus. Our findings underline the importance of correct result interpretation and positivity cut-off set-up for different detection methods of A. phagocytophilum. This bacterium is characterized by a high intraspecific variability and highly sensitive detection itself, is not enough. Detailed molecular typing is necessary to define the zoonotic potential of different strains and their natural reservoirs.

• MeSH
• Anaplasma phagocytophilum * genetika   MeSH
• fylogeneze MeSH
• klíště * mikrobiologie   MeSH
• zajíci * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Evropa MeSH

Extracellular vesicles are thought to facilitate pathogen transmission from arthropods to humans and other animals. Here, we reveal that pathogen spreading from arthropods to the mammalian host is multifaceted. Extracellular vesicles from Ixodes scapularis enable tick feeding and promote infection of the mildly virulent rickettsial agent Anaplasma phagocytophilum through the SNARE proteins Vamp33 and Synaptobrevin 2 and dendritic epidermal T cells. However, extracellular vesicles from the tick Dermacentor andersoni mitigate microbial spreading caused by the lethal pathogen Francisella tularensis. Collectively, we establish that tick extracellular vesicles foster distinct outcomes of bacterial infection and assist in vector feeding by acting on skin immunity. Thus, the biology of arthropods should be taken into consideration when developing strategies to control vector-borne diseases.

• MeSH
• Anaplasma phagocytophilum patogenita   MeSH
• bakteriální infekce imunologie   metabolismus   MeSH
• buněčné linie MeSH
• členovci metabolismus   mikrobiologie   fyziologie   MeSH
• Dermacentor metabolismus   mikrobiologie   fyziologie   MeSH
• extracelulární vezikuly metabolismus   ultrastruktura   MeSH
• Francisella tularensis patogenita   MeSH
• genová ontologie MeSH
• intravitální mikroskopie MeSH
• klíšťata metabolismus   mikrobiologie   MeSH
• klíště metabolismus   mikrobiologie   fyziologie   MeSH
• kůže imunologie   mikrobiologie   parazitologie   MeSH
• lidé MeSH
• membránový protein 2 asociovaný s vezikuly metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši MeSH
• proteiny R-SNARE metabolismus   MeSH
• proteomika MeSH
• T-lymfocyty metabolismus   MeSH
• tandemová hmotnostní spektrometrie MeSH
• transmisní elektronová mikroskopie MeSH
• zánět imunologie   metabolismus   parazitologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH

BACKGROUND: Phylogenetically closely related strains of maternally inherited endosymbiotic bacteria are often found in phylogenetically divergent, and geographically distant insect host species. The interspecies transfer of the symbiont Wolbachia has been thought to have occurred repeatedly, facilitating its observed global pandemic. Few ecological interactions have been proposed as potential routes for the horizontal transfer of Wolbachia within natural insect communities. These routes are however likely to act only at the local scale, but how they may support the global distribution of some Wolbachia strains remains unclear. RESULTS: Here, we characterize the Wolbachia diversity in butterflies from the tropical forest regions of central Africa to discuss transfer at both local and global scales. We show that numerous species from both the Mylothris (family Pieridae) and Bicyclus (family Nymphalidae) butterfly genera are infected with similar Wolbachia strains, despite only minor interclade contacts across the life cycles of the species within their partially overlapping ecological niches. The phylogenetic distance and differences in resource use between these genera rule out the role of ancestry, hybridization, and shared host-plants in the interspecies transfer of the symbiont. Furthermore, we could not identify any shared ecological factors to explain the presence of the strains in other arthropod species from other habitats, or even ecoregions. CONCLUSION: Only the systematic surveys of the Wolbachia strains from entire species communities may offer the material currently lacking for understanding how Wolbachia may transfer between highly different and unrelated hosts, as well as across environmental scales.

• MeSH
• biodiverzita MeSH
• fylogeneze MeSH
• genom bakteriální MeSH
• Lepidoptera genetika   fyziologie   MeSH
• přenos genů horizontální * MeSH
• symbióza * MeSH
• Wolbachia genetika   fyziologie   MeSH
• zeměpis MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Afrika 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