(1) Background: Ixodes ricinus is responsible for the spreading of medically important pathogens. Monitoring the level of tick infection in various areas is essential for determining the potential tick-born risk. This study aimed to detect Borrelia spp. and Rickettsia spp. in I. ricinus ticks collected in urban and protected areas both in Poland and the Czech Republic. (2) Methods: Ticks were collected by flagging in the years 2016-2017. Borrelia spp. was detected using nested PCR targeting the flaB gene and Rickettsia spp. using nested PCR targeting gltA. (3) Results: In total, DNA of Borrelia spp. was detected in 25.9% of samples. Ticks collected in Poland were more infected compared to the Czech Republic and ticks collected in protected areas were more infected with Borrelia spp. than ticks collected in urban areas. The RFLP analysis showed the occurrence of B. afzelii and B. garinii in both countries, and additionally B. valaisiana, B. burgdorferi s.s., and B. miyamotoi in Poland. Rickettsia spp. was detected in 17.4% of I. ricinus, with comparable infection level in both countries; however, regional differences were observed. (4) Conclusion: The regional differences in Borrelia spp. and Rickettsia spp. prevalence in I. ricinus indicate the complexity of factors influencing the level of infection and underline the need for adaptation public health surveillance strategies in each region.

• Keywords
• Borrelia spp., Ixodes ricinus, Rickettsia spp., protected areas, tick-borne risk, urban areas,
• Publication type
• Journal Article MeSH

Wild small mammals and ticks play an important role in maintaining and spreading zoonoses in nature, as well as in captive animals. The aim of this study was to monitor selected agents with zoonotic potential in their reservoirs and vectors in a zoo, and to draw attention to the risk of possible contact with these pathogens. In total, 117 wild small mammals (rodents) and 166 ticks were collected in the area of Brno Zoo. Antibodies to the bacteria Coxiella burnetii, Francisella tularensis, and Borrelia burgdorferi s.l. were detected by a modified enzyme-linked immunosorbent assay in 19% (19/99), 4% (4/99), and 15% (15/99) of rodents, respectively. Antibodies to Leptospira spp. bacteria were detected by the microscopic agglutination test in 6% (4/63) of rodents. Coinfection (antibodies to more than two agents) were proved in 14.5% (15/97) of animals. The prevalence of C. burnetii statistically differed according to the years of trapping (p = 0.0241). The DNAs of B. burgdorferi s.l., Rickettsia sp., and Anaplasma phagocytophilum were detected by PCR in 16%, 6%, and 1% of ticks, respectively, without coinfection and without effect of life stage and sex of ticks on positivity. Sequencing showed homology with R. helvetica and A. phagocytophilum in four and one positive samples, respectively. The results of our study show that wild small mammals and ticks in a zoo could serve as reservoirs and vectors of infectious agents with zoonotic potential and thus present a risk of infection to zoo animals and also to keepers and visitors to a zoo.

• Keywords
• Anaplasma, Borrelia, Coxiella, Francisella, Leptospira, Rickettsia,
• Publication type
• Journal Article 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.

• Keywords
• Anaplasma phagocytophilum, Ixodes ricinus, deer, ecotype,
• MeSH
• Anaplasma phagocytophilum genetics   isolation & purification   MeSH
• Ecosystem MeSH
• Ecotype MeSH
• Ixodes genetics   microbiology   MeSH
• Humans MeSH
• RNA, Ribosomal, 16S genetics   MeSH
• Deer microbiology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Europe MeSH
• Names of Substances
• RNA, Ribosomal, 16S MeSH

BACKGROUND: Candidatus Neoehrlichia mikurensis (CNM) is an emerging tick-borne pathogen causing severe disease in immunocompromised patients. In Europe, Ixodes ricinus is the primary vector and rodents act as reservoir hosts. New data on the prevalence of CNM in ticks and rodents contribute to the knowledge on the distribution of endemic areas and circulation of the bacterium in natural foci. METHODS: Questing ticks were collected and rodents were trapped in urban/suburban and natural habitats in South-Western Slovakia from 2011 to 2014. DNA from questing and rodent-attached ticks and rodent tissues were screened for CNM by real-time PCR. Rodent spleen samples positive for CNM were characterised at the groEL gene locus. Spatial and temporal differences in CNM prevalence in ticks and rodents and co-infections of ticks with CNM and Anaplasma phagocytophilum were analysed. RESULTS: The presence of CNM was confirmed in questing and rodent-attached I. ricinus ticks and in rodents. Total prevalence in both ticks and rodents was significantly higher in the natural habitat (2.3% and 10.1%, respectively) than in the urban/suburban habitat (1.0% and 3.3%, respectively). No seasonal pattern in CNM prevalence in ticks was observed, but prevalence in rodents was higher in autumn than in spring. CNM was detected in Apodemus flavicollis, Myodes glareolus, Microtus arvalis and Micromys minutus, with the highest prevalence in M. arvalis (30%). By screening CNM dissemination in rodent tissues, infection was detected in lungs of all specimens with positive spleens and in blood, kidney, liver and skin of part of those individuals. Infection with CNM was detected in 1.3% of rodent attached I. ricinus ticks. Sequences of a fragment of the groEL gene from CNM-positive rodents showed a high degree of identity with sequences of the gene amplified from ticks and infected human blood from Europe. Only 0.1% of CNM-positive questing ticks carried A. phagocytophilum. Ticks infected with CNM prevailed in the natural habitat (67.2%), whereas ticks infected with A. phagocytophilum prevailed in the urban/suburban habitat (75.0%). CONCLUSION: The study confirmed the circulation of CNM between I. ricinus ticks and rodents in South-Western Slovakia, and indicates a potential risk of contracting human infections.

• MeSH
• Anaplasma phagocytophilum genetics   isolation & purification   MeSH
• Anaplasmataceae genetics   isolation & purification   MeSH
• Arachnid Vectors microbiology   MeSH
• Ehrlichiosis epidemiology   microbiology   MeSH
• Ecosystem MeSH
• Rodentia MeSH
• Anaplasmataceae Infections epidemiology   microbiology   MeSH
• Tick Infestations epidemiology   microbiology   MeSH
• Ixodes microbiology   MeSH
• Coinfection MeSH
• Humans MeSH
• Risk MeSH
• Disease Reservoirs microbiology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Slovakia epidemiology   MeSH

Bacteria associated with the tick Ixodes ricinus were assessed in specimens unattached or attached to the skin of cats, dogs and humans, collected in the Czech Republic. The bacteria were detected by PCR in 97 of 142 pooled samples including 204 ticks, i.e. 1-7 ticks per sample, collected at the same time from one host. A fragment of the bacterial 16S rRNA gene was amplified, cloned and sequenced from 32 randomly selected samples. The most frequent sequences were those related to Candidatus Midichloria midichlori (71% of cloned sequences), followed by Diplorickettsia (13%), Spiroplasma (3%), Rickettsia (3%), Pasteurella (3%), Morganella (3%), Pseudomonas (2%), Bacillus (1%), Methylobacterium (1%) and Phyllobacterium (1%). The phylogenetic analysis of Spiroplasma 16S rRNA gene sequences showed two groups related to Spiroplasma eriocheiris and Spiroplasma melliferum, respectively. Using group-specific primers, the following potentially pathogenic bacteria were detected: Borellia (in 20% of the 142 samples), Rickettsia (12%), Spiroplasma (5%), Diplorickettsia (5%) and Anaplasma (2%). In total, 68% of I. ricinus samples (97/142) contained detectable bacteria and 13% contained two or more putative pathogenic groups. The prevalence of tick-borne bacteria was similar to the observations in other European countries.

• Keywords
• Bacteria, Central Bohemia, Ixodes ricinus, Pathogens, Tick,
• MeSH
• Bacteria classification   genetics   MeSH
• DNA, Bacterial genetics   MeSH
• Phylogeny MeSH
• Ixodes growth & development   microbiology   MeSH
• Cats microbiology   parasitology   MeSH
• Humans MeSH
• Molecular Sequence Data MeSH
• Nymph microbiology   MeSH
• Polymerase Chain Reaction MeSH
• Dogs microbiology   parasitology   MeSH
• RNA, Ribosomal, 16S genetics   MeSH
• Spiroplasma classification   genetics   MeSH
• Cities MeSH
• Animals MeSH
• Check Tag
• Cats microbiology   parasitology   MeSH
• Humans MeSH
• Male MeSH
• Dogs microbiology   parasitology   MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Czech Republic MeSH
• Cities MeSH
• Names of Substances
• DNA, Bacterial MeSH
• RNA, Ribosomal, 16S MeSH

BACKGROUND: Ixodes ricinus is the principal vector of Anaplasma phagocytophilum, the ethiological agent of granulocytic anaplasmosis in Europe. Anaplasmosis is an emerging zoonotic disease with a natural enzootic cycle. The reservoir competence of rodents is unclear. Monitoring of A. phagocytophilum prevalence in I. ricinus and rodents in various habitat types of Slovakia may contribute to the knowledge about the epidemiology of anaplasmosis in Central Europe. METHODS: Over 4400 questing ixodid ticks, 1000 rodent-attached ticks and tissue samples of 606 rodents were screened for A. phagocytophilum DNA by real-time PCR targeting the msp2 gene. Ticks and rodents were captured along six transects in an urban/suburban and natural habitat in south-western Slovakia during 2011-2014. Estimates of wildlife (roe deer, red deer, fallow deer, mouflon, wild boar) densities in the study area were taken from hunter's yearly reports. Spatial and temporal differences in A. phagocytophilum prevalence in questing I. ricinus and relationships with relative abundance of ticks and wildlife were analysed. RESULTS: Overall prevalence of A. phagocytophilum in questing I. ricinus was significantly higher in the urban/suburban habitat (7.2%; 95% CI: 6.1-8.3%) compared to the natural habitat (3.1%; 95% CI: 2.5-3.9%) (χ(2) = 37.451; P < 0.001). Significant local differences in prevalence of infected questing ticks were found among transects within each habitat as well as among years and between seasons. The trapped rodents belonged to six species. Apodemus flavicollis and Myodes glareolus prevailed in both habitats, Microtus arvalis was present only in the natural habitat. I. ricinus comprised 96.3% of the rodent-attached ticks, the rest were Haemaphysalis concinna, Ixodes trianguliceps and Dermacentor reticulatus. Only 0.5% of rodent skin and 0.6% of rodent-attached ticks (only I. ricinus) were infected with A. phagocytophilum. Prevalence of A. phagocytophilum in questing I. ricinus did not correlate significantly with relative abundance of ticks or with abundance of wildlife in the area. CONCLUSION: The study confirms that urban I. ricinus populations are infected with A. phagocytophilum at a higher rate than in a natural habitat of south-western Slovakia and suggests that rodents are not the main reservoirs of the bacterium in the investigated area.

• MeSH
• Anaplasma phagocytophilum isolation & purification   MeSH
• Time Factors MeSH
• Ehrlichiosis epidemiology   transmission   MeSH
• Ecosystem MeSH
• Rodentia MeSH
• Ixodes microbiology   MeSH
• Real-Time Polymerase Chain Reaction MeSH
• Rodent Diseases epidemiology   microbiology   transmission   MeSH
• Nymph microbiology   MeSH
• Animal Distribution MeSH
• Cities MeSH
• Disease Reservoirs * microbiology   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Slovakia epidemiology   MeSH
• Cities MeSH

Tick-borne diseases represent major public and animal health issues worldwide. Ixodes ricinus, primarily associated with deciduous and mixed forests, is the principal vector of causative agents of viral, bacterial, and protozoan zoonotic diseases in Europe. Recently, abundant tick populations have been observed in European urban green areas, which are of public health relevance due to the exposure of humans and domesticated animals to potentially infected ticks. In urban habitats, small and medium-sized mammals, birds, companion animals (dogs and cats), and larger mammals (roe deer and wild boar) play a role in maintenance of tick populations and as reservoirs of tick-borne pathogens. Presence of ticks infected with tick-borne encephalitis virus and high prevalence of ticks infected with Borrelia burgdorferi s.l., causing Lyme borreliosis, have been reported from urbanized areas in Europe. Emerging pathogens, including bacteria of the order Rickettsiales (Anaplasma phagocytophilum, "Candidatus Neoehrlichia mikurensis," Rickettsia helvetica, and R. monacensis), Borrelia miyamotoi, and protozoans (Babesia divergens, B. venatorum, and B. microti) have also been detected in urban tick populations. Understanding the ecology of ticks and their associations with hosts in a European urbanized environment is crucial to quantify parameters necessary for risk pre-assessment and identification of public health strategies for control and prevention of tick-borne diseases.

• Keywords
• Europe, Ixodes ricinus, tick-borne pathogens, ticks, urban habitats,
• Publication type
• Journal Article MeSH
• Review MeSH