Celkem jsme v roce 2007 vyšetřili 1048 jedinců klíštěte Ixodes ricinus z toho 935 ze sběrů vlajkováním v pražských parcích a 114 jedinců ze zákusu na člověku. V roce 2008 jsme vyšetřili 364 jedinců z toho 277 ze sběru a 86 sejmutých z lidí. Promořenost klíšťat původcem lymeské borreliózy v roce 2007 byla 9,7 %, zjištěná v temném poli a v 13 % byla prokázána DNA pomocí PCR. V roce 2008 byla promořenost klíšťat vyšší, 14 % v temném poli a 20 % pomocí PCR. Klíšťata sejmutá po zákusu na lidech byla infikována v 7 %. Přítomnost Anaplasma phagocytophilum, Bartonella sp. a Rickettsia sp byla prokázána pomocí polymerázové řetězové reakce a kontrolována sekvencí geonomu u 11,3 - 11,8 % klíšťat. Spiroplasma rodu Babesia byla zjištěna v 7,4 % u samic I. ricinus. Klíšťata sejmutá z lidí byla infikovaná borrelií v 6,5 % a anaplasmou v 1,5 %. Pravidelnými sběry v jednotlivých měsících r. 2007 a 2008 na jedné lokalitě v Praze 10 jsme ověřili změny výskytu ruzných stádií klíštěte a jejich měnící se nákazu ruznými agens v závislosti na teplotě a vývoji. K růstu bakterií Borrelia a Anaplasma sp. ve střevě klíštěte je zapotřebí určité doby s vyšší teplotou. Nejmenší počet infikovaných klíšťat byl v srpnu 2007 a 2008, kdy se z velké většiny vyskytovaly pouze larvy.

In 2007, 1048 Ixodes ricinus ticks were investigated: 935 of these were collected by flagging in Prague parks and 113 were ticks attached to the human skin. In 2008, 364 ticks were investigated, i.e. 277 and 87 ticks collected by flagging and from humans, respectively. In 2007, the causative agent of lyme borreliosis was detected in 9.7% of ticks in dark field and in 13% of ticks in DNA by PCR. In 2008, higher positivity rates were found, i.e. 14% and 20%, respectively. Seven percent of ticks obtained from humans were infected. Anaplasma phagocytophilum, Bartonella sp. and Rickettsia sp. were detected by PCR and sequence analysis in 11.3% - 11.8% of ticks. Spiroplasma of the genus Babesia was detected in 7.4% of I. ricinus females. The ticks collected from humans were infected by Borrelia in 6.5% and by Anaplasma in 1.5%. Regular monthly flagging was performed in one locality in Prague 10 in 2007 and 2008 to obtain data on the incidence of different stages of ticks and rates of infection by different agents depending on temperature and season. To grow in the tick intestine, Borrelia and Anaplasma need higher temperature for a certain period of time. The lowest numbers of infected ticks were found in August 2007 and 2008 when mostly tick larvae were collected.

• MeSH
• Anaplasma phagocytophilum genetics   isolation & purification   MeSH
• Babesia genetics   isolation & purification   MeSH
• Bartonella genetics   isolation & purification   MeSH
• Borrelia burgdorferi genetics   isolation & purification   MeSH
• Disease Vectors MeSH
• Ixodes microbiology   MeSH
• Lyme Disease epidemiology   transmission   MeSH
• Polymerase Chain Reaction utilization   MeSH
• Publication type
• Chart 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

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 isolation & purification   MeSH
• Ctenocephalides microbiology   MeSH
• Rickettsia isolation & purification   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Poland MeSH

Cíl: Zjistit výskyt potenciálně patogenních druhů babesií pro člověka v klíšťatech a v krvi psů a jelenů ve vybraných regionech České republiky. Prevalenci Babesia spp. v klíšťatech porovnat s výskytem jiných patogenů přenášených klíšťaty jako Borrelia spp., Anaplasma spp., Rickettsia spp. Materiál a metody: Vzorky klíšťat byly jednotlivě homogenizovány, ze vzorků klíšťat a krve živočichů provedena izolace DNA. Detekce Babesia spp. byla stanovena metodou PCR 18S rRNA genu a sekvenační analýzou PCR produktů určeny jednotlivé druhy babesií. Výsledky: V letech 2014–2016 byla analyzována klíšťata a krev psů a jelenů na různých místech České republiky. Ze souboru 675 klíšťat Ixodes ricinus dosahovala pozitivita na přítomnost Babesia spp. hodnot od 0,0 do 3,3 %. Sekvenační analýzou byly v klíšťatech identifikovány druhy Babesia venatorum, Babesia microti (patogenní druhy pro člověka) a druh Babesia capreoli. Prevalence Babesia spp. v klíšťatech byla v porovnání s výskytem jiných patogenů jako Borrelia burgdorferi s. l. (29,3 %), Anaplasma phagocytophilum (4,9 %) nižší a srovnatelná s Rickettsia spp. (1,6 %). U třetiny pozitivních klíšťat na babesie byla zjištěna koinfekce s Borrelia burgdorferi s. l. (B. venatorum – Borrelia garinii, Borrelia afzelii a B. microti – B. afzelii). Ze 109 vzorků krve psů bylo 3,7 % pozitivních na Babesia spp. s výskytem druhů Babesia gibsoni a Babesia vulpes. Z 50 vzorků krve jelenů z přírodního ekosystému dosahovala pozitivita 4,0 %. Identifikován byl druh Babesia divergens, nejvíce patogenní druh Babesia spp. pro člověka. Z 80 vzorků krve jelenů chovaných na farmách bylo pozitivních 5,0 % s výskytem druhu Babesia odocoilei. Nukleotidové sekvence babesií způsobujících humánní babesiózu byly zaslány do genové banky a přijaty pod čísly ON892053 (B. venatorum), ON892061 (B. microti), ON892067 (B. divergens). Závěr: Metodou PCR 18S rRNA genu a sekvenací amplikonů byly na území České republiky detekovány tři druhy babesií patogenních pro člověka: B. divergens, B. venatorum, B. microti. Výskyt těchto druhů babesií znamená potenciální riziko onemocnění babesiózou, zejména pro asplenické a imunokompromitované pacienty. Zjištěné koinfekce s Borrelia burgdorferi s. l. mohou být příčinou komplikovaného průběhu onemocnění.

Aim: To determine the occurrence of species of Babesia potentially pathogenic for humans in ticks and in the blood of dogs and deer in selected regions of the Czech Republic. To compare the prevalence of Babesia spp. in ticks with that of other tick-borne pathogens, such as Borrelia spp., Anaplasma spp., and Rickettsia spp. Material and Methods: Tick samples were individually homogenized. DNA was isolated from tick samples and animal blood. The detection of Babesia spp. was based on PCR of the 18S rRNA gene, and the identification to the species level was done by sequencing analysis of the PCR products. Results: In 2014–2016, ticks and blood of dogs and deer collected in various areas of the Czech Republic were analyzed. In a set of 675 Ixodes ricinus ticks, the positivity rate for Babesia spp. varied from 0.0 to 3.3 %. The species Babesia venatorum, Babesia microti (both pathogenic for humans), and Babesia capreoli were identified in ticks by sequencing analysis. The prevalence of Babesia spp. in ticks compared to that of other pathogens such as Borrelia burgdorferi s. l. (29.3 %) or Anaplasma phagocytophilum (4.9 %) was lower and comparable to that of Rickettsia spp. (1.6 %). Co-infection with Borrelia burgdorferi s.l (B. venatorum – Borrelia garinii, Borrelia afzelii, and B. microti – B. afzelii) was found in a third of Babesia spp. positive ticks. Out of 109 dog blood samples, 3.7 % were positive for Babesia spp., specifically Babesia gibsoni and Babesia vulpes. Of 50 blood samples of wild deer from the natural ecosystem, the positivity rate reached 4.0 %. The species Babesia divergens, a major human pathogen, was identified. Out of 80 blood samples from farmed deer, 5.0 % were positive for the species Babesia odocoilei. Nucleotide sequences of the agents causing human babesiosis were deposited in the gene bank under accession numbers ON892053 (B. venatorum), ON892061 (B. microti), and ON892067 (B. divergens). Conclusions: Using PCR of the 18S rRNA gene and amplicon sequencing, three species of Babesia causing human babesiosis were detected in the Czech Republic: B. divergens, B. venatorum, and B. microti. Babesia spp. pathogenic for humans pose a potential risk especially in asplenic and immunocompromised patients. The detected co-infections with Borrelia spp. can be the cause of a complicated course of the disease.

• MeSH
• Babesia microbiology   MeSH
• Babesiosis * epidemiology   blood   transmission   MeSH
• Borrelia burgdorferi MeSH
• Molecular Diagnostic Techniques methods   MeSH
• Ticks * microbiology   MeSH
• Coinfection diagnosis   transmission   MeSH
• Blood microbiology   MeSH
• Humans MeSH
• Tick-Borne Diseases epidemiology   transmission   prevention & control   MeSH
• Polymerase Chain Reaction methods   MeSH
• Dogs * microbiology   MeSH
• Deer * blood   microbiology   MeSH
• Check Tag
• Humans MeSH
• Dogs * microbiology   MeSH
• Geographicals
• Czech Republic MeSH

Wild birds are known to be a reservoir of infectious disease agents and disseminatory hosts of ticks. The purpose of this work was to obtain information about the occurrence of rickettsial, anaplasmal, and borrelial infections in some ticks that parasitize wild birds in the Czech Republic. A total of 549 subadult ticks of three species Ixodes arboricola (75.0%), Ixodes ricinus (23.1%), and Haemaphysalis concinna (1.8%) were collected from 20 species of birds (Passeriformes). Rickettsiae were detected in 44.0% larvae and 24.5% nymphs of I. arboricola collected from Parus major, Poecile palustris, and Sitta europaea. Rickettsiae-positive I. ricinus larvae (13.7%) were collected from P. major, Cyanistes caeruleus, and S. europaea, and 2.6% of nymphs from Erithacus rubecula and Prunella modularis. Comparison of sequences of a gltA gene fragment with data available in GenBank identified Rickettsia helvetica, a spotted fever rickettsia associated with human infections, and other Rickettsia spp. Anaplasma phagocytophilum was found only in two I. ricinus nymphs collected from E. rubecula and P. major. Infections with Borrelia burgdorferi sensu lato were recorded in 1.3% larvae of I. arboricola acquired from P. palustris and P. major and in 11.8% larvae and 25.0% nymphs of I. ricinus collected from P. major, P. palustris, C. caeruleus, Acrocephalus schoenobaenus, Turdus merula, Carpodacus erythrinus, Sylvia atricapilla, P. modularis, and Phylloscopus collybita. Reverse-line blot hybridization showed infections with Borrelia garinii and Borrelia valaisiana and mixed infections with these two genospecies. This is the first record of a high rate of rickettsial infection in I. arboricola subadult ticks acquired from birds in the Czech Republic and in central Europe. Our study suggests that I. arboricola, P. major, and P. palustris play important roles in circulating rickettsiae.

• MeSH
• Anaplasma phagocytophilum isolation & purification   MeSH
• Arachnid Vectors microbiology   MeSH
• Borrelia burgdorferi isolation & purification   MeSH
• Databases, Nucleic Acid MeSH
• Ixodes microbiology   MeSH
• Larva microbiology   MeSH
• Nymph microbiology   MeSH
• Polymerase Chain Reaction MeSH
• Birds parasitology   MeSH
• Rickettsia genetics   isolation & purification   MeSH
• Sequence Analysis MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Czech Republic MeSH

Distribution of Anaplasma spp., Babesia spp., Theileria spp., and Ehrlichia ruminantium, was for the first time studied in Bié Province, central Angola. We examined 76 blood samples of cattle originated from seven farms, and 13 blood samples of goats from two farms employing molecular genetic tools (PCR). Most prevalent was A. ovis-infection in goats (100%) and A. marginale-infection in cattle (38% of examined animals, and six out of seven farms). B. bigemina-infection was detected in only one specimen at Andulo, whereas B. bovis was not detected in Bié. We did not detected T. parva, the causative agent of serious diseases in cattle; nevertheless, infection by T. velifera was quite frequent (14% of examined animals, and five out of seven farms). Causative agent of heartwater disease - E. ruminantium, was not detected. Taking into account short-term perspective of PCR methods in monitoring of epidemiological status in herds, the number of infected animals and distribution of detected pathogens should not be ignored.

• MeSH
• Anaplasma genetics   isolation & purification   MeSH
• Anaplasmosis epidemiology   microbiology   MeSH
• Babesia genetics   isolation & purification   MeSH
• Babesiosis epidemiology   parasitology   veterinary   MeSH
• DNA, Bacterial blood   chemistry   isolation & purification   MeSH
• Goats MeSH
• Goat Diseases epidemiology   microbiology   parasitology   MeSH
• Tick-Borne Diseases epidemiology   microbiology   parasitology   veterinary   MeSH
• Cattle Diseases epidemiology   microbiology   parasitology   MeSH
• Polymerase Chain Reaction veterinary   MeSH
• DNA, Protozoan blood   chemistry   isolation & purification   MeSH
• Base Sequence MeSH
• Cattle MeSH
• Theileria genetics   isolation & purification   MeSH
• Theileriasis epidemiology   parasitology   MeSH
• Animals MeSH
• Check Tag
• Cattle MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Angola 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 * classification   genetics   isolation & purification   MeSH
• Babesia genetics   MeSH
• Bartonella genetics   MeSH
• DNA, Bacterial chemistry   genetics   isolation & purification   MeSH
• Ehrlichiosis complications   epidemiology   pathology   veterinary   MeSH
• Phylogeny MeSH
• Oligonucleotides chemistry   MeSH
• Polymerase Chain Reaction veterinary   MeSH
• Base Sequence MeSH
• Spleen microbiology   pathology   MeSH
• Body Weight MeSH
• Deer * MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Norway MeSH

Anaplasma phagocytophilum has been first isolated from the blood of two Czech patients simultaneously with a cultivation of Borrelia burgdorferi sensu lato from their erythema migrans lesions. Cultivation of different Borrelia spp. from 12 erythema migrans biopsies, from 2 blood, one liquor and one placenta sample in BSK-H medium was successful. Adapted conventional methods targeting 16S rRNA and OspA genes for real-time polymerase chain reaction (PCR) and partial sequencing of these genes together with microscopical examinations of the blood smears provided a direct detection of the B. afzelii, B. burgdorferi, B. garinii, B. valaisiana and B. bissettii in the skin, B. garinii in the blood, placenta and liquor in 24 (36.3 %) patients, and A. phagocytophilum in 10 (15 %) patients with erythema migrans. Positive indirect IgM immunofluorescence against Anaplasma sp. was obtained in 7 cases, specific IgG antibodies were detected in 12 patients. Three women suffering from erythema migrans in the first trimester had positive PCR for Anaplasma and/or for Borrelia in the blood and two of them, later, in the placenta. Interpretation of laboratory data can bring important contribution to establishing the role of Anaplasma sp. in erythema migrans and forming the principle of precaution with laboratory diagnosis during pregnancy which always should be reflected in the resistance of Anaplasma sp. toward penicillins.

• MeSH
• Anaplasma phagocytophilum immunology   isolation & purification   MeSH
• Antigens, Surface genetics   MeSH
• Bacterial Vaccines genetics   MeSH
• Borrelia burgdorferi genetics   immunology   isolation & purification   MeSH
• Diagnosis, Differential MeSH
• Adult MeSH
• Ehrlichiosis diagnosis   blood   microbiology   MeSH
• Erythema Chronicum Migrans diagnosis   blood   microbiology   MeSH
• Financing, Organized MeSH
• HL-60 Cells MeSH
• Pregnancy Complications, Infectious diagnosis   microbiology   MeSH
• Middle Aged MeSH
• Humans MeSH
• Lipoproteins genetics   MeSH
• Lyme Disease diagnosis   blood   microbiology   MeSH
• Adolescent MeSH
• Placenta microbiology   MeSH
• Bacterial Outer Membrane Proteins genetics   MeSH
• Antibodies, Bacterial blood   MeSH
• Aged MeSH
• Pregnancy MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Adolescent MeSH
• Male MeSH
• Aged MeSH
• Pregnancy MeSH
• Female MeSH

• Publication type
• Abstracts MeSH

Ticks are obligate hematophagous arthropods of significant importance to human and veterinary medicine. They transmit a vast array of pathogens, including bacteria, viruses, protozoa, and helminths. Most epidemiological data on ticks and tick-borne pathogens (TBPs) in the West Indies are limited to common livestock pathogens such as Ehrlichia ruminantium, Babesia spp. (i.e., B. bovis and B. bigemina), and Anaplasma marginale, and less information is available on companion animal pathogens. Of note, human tick-borne diseases (TBDs) remain almost completely uncharacterized in the West Indies. Information on TBP presence in wildlife is also missing. Herein, we provide a comprehensive review of the ticks and TBPs affecting human and animal health in the Caribbean, and introduce the challenges associated with understanding TBD epidemiology and implementing successful TBD management in this region. In particular, we stress the need for innovative and versatile surveillance tools using high-throughput pathogen detection (e.g., high-throughput real-time microfluidic PCR). The use of such tools in large epidemiological surveys will likely improve TBD prevention and control programs in the Caribbean.

• MeSH
• Anaplasma marginale isolation & purification   pathogenicity   MeSH
• Babesia isolation & purification   pathogenicity   MeSH
• Animals, Wild MeSH
• Ehrlichia ruminantium isolation & purification   pathogenicity   MeSH
• Epidemiological Monitoring veterinary   MeSH
• Insect Vectors classification   microbiology   parasitology   MeSH
• Ticks classification   microbiology   parasitology   MeSH
• Humans MeSH
• Tick-Borne Diseases epidemiology   microbiology   parasitology   prevention & control   MeSH
• Animal Diseases epidemiology   microbiology   parasitology   MeSH
• High-Throughput Screening Assays methods   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH
• Geographicals
• Caribbean Region epidemiology   MeSH
• West Indies epidemiology   MeSH