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 * izolace a purifikace   MeSH
• Borrelia izolace a purifikace   MeSH
• Coxiella izolace a purifikace   genetika   MeSH
• infestace klíšťaty veterinární   epidemiologie   parazitologie   MeSH
• Ixodidae * mikrobiologie   růst a vývoj   MeSH
• Rickettsia * izolace a purifikace   MeSH
• želvy * mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Střední východ MeSH
• Středomoří 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 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

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

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

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 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

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

The way in which European genetic variants of Anaplasma phagocytophilum circulate in their natural foci and which variants cause disease in humans or livestock remains thus far unclear. Red deer and roe deer are suggested to be reservoirs for some European A. phagocytophilum strains, and Ixodes ricinus is their principal vector. Based on groEL gene sequences, five A. phagocytophilum ecotypes have been identified. Ecotype I is associated with the broadest host range, including strains that cause disease in domestic animals and humans. Ecotype II is associated with roe deer and does not include zoonotic strains. In the present study, questing I. ricinus were collected in urban, pasture, and natural habitats in the Czech Republic, Germany, and Slovakia. A fragment of the msp2 gene of A. phagocytophilum was amplified by real-time PCR in DNA isolated from ticks. Positive samples were further analyzed by nested PCRs targeting fragments of the 16S rRNA and groEL genes, followed by sequencing. Samples were stratified according to the presence/absence of roe deer at the sampling sites. Geographic origin, habitat, and tick stage were also considered. The probability that A. phagocytophilum is a particular ecotype was estimated by a generalized linear model. Anaplasma phagocytophilum was identified by genetic typing in 274 I. ricinus ticks. The majority belonged to ecotype I (63.9%), 28.5% were ecotype II, and both ecotypes were identified in 7.7% of ticks. Ecotype II was more frequently identified in ticks originating from a site with presence of roe deer, whereas ecotype I was more frequent in adult ticks than in nymphs. Models taking into account the country-specific, site-specific, and habitat-specific aspects did not improve the goodness of the fit. Thus, roe deer presence in a certain site and the tick developmental stage are suggested to be the two factors consistently influencing the occurrence of a particular A. phagocytophilum ecotype in a positive I. ricinus tick.

• MeSH
• Anaplasma phagocytophilum genetika   izolace a purifikace   MeSH
• ekosystém MeSH
• ekotyp MeSH
• klíště genetika   mikrobiologie   MeSH
• lidé MeSH
• RNA ribozomální 16S genetika   MeSH
• vysoká zvěř mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Evropa 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