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Isolation, cultivation and identification of Borrelia burgdorferi genospecies from Ixodes ricinus ticks from the city of Brno, Czech Republic
Pejchalová K, Zákovská A, Mejzlíková M, Halouzka J, Dendis M
Language English Country Poland
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- MeSH
- Arachnid Vectors microbiology MeSH
- Borrelia burgdorferi Group genetics isolation & purification classification MeSH
- DNA, Bacterial isolation & purification MeSH
- Species Specificity MeSH
- Financing, Organized MeSH
- Phylogeny MeSH
- Ixodes microbiology MeSH
- Larva microbiology MeSH
- Humans MeSH
- Lyme Disease epidemiology transmission MeSH
- Environmental Monitoring MeSH
- Nymph microbiology MeSH
- Polymerase Chain Reaction methods MeSH
- Polymorphism, Restriction Fragment Length MeSH
- Prevalence MeSH
- Risk Factors MeSH
- Disease Reservoirs microbiology veterinary MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Female MeSH
- Animals MeSH
- Geographicals
- Czech Republic MeSH
A total of 305 ticks (21 larvae, 243 nymphs, 19 females and 22 males) were collected by flagging of vegetation in suburban woods of Pisarky Park (city of Brno) from July to October 2002. The midgut of each tick was dissected out and transferred individually into BSK-H medium. After cultivation, all specimens were examined by dark-field microscopy (DFM) for the presence of borreliae. Out of 305 tick samples, 45 were (14.8%) DFM positive. The following polymerase chain reaction (PCR) then revealed 37 (12.1%) samples positive for the presence of Borrelia burgdorferi sensu lato DNA. All 37 samples were further analysed by restriction fragment length polymorphism (RFLP) method. PCR-RFLP analysis revealed 14 strains of B. afzelii (37.8%), 15 strains of B. garinii (40.5%) and 2 strains of B. burgdorferi sensu stricto (5.4%). Four samples (10.8%) showed a mixed population of these genospecies. Two samples produced atypical RFLP pattern which were detected by sequence analysis as B. valaisiana (5.4%). Isolation attempts resulted in 21 spirochaetal strains (including two stains of B. valaisiana). The results show the diversity of B. burgdorferi s.l. in tick population and refer the risk of infection by pathogenic borreliae in Brno.
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- $a Department of Anatomy, Division of Neuroanatomy, Faculty of Medicine, Masaryk University, Kamenice 3, 625 00 Brno, Czech Republic. keith@sci.muni.cz
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- $a A total of 305 ticks (21 larvae, 243 nymphs, 19 females and 22 males) were collected by flagging of vegetation in suburban woods of Pisarky Park (city of Brno) from July to October 2002. The midgut of each tick was dissected out and transferred individually into BSK-H medium. After cultivation, all specimens were examined by dark-field microscopy (DFM) for the presence of borreliae. Out of 305 tick samples, 45 were (14.8%) DFM positive. The following polymerase chain reaction (PCR) then revealed 37 (12.1%) samples positive for the presence of Borrelia burgdorferi sensu lato DNA. All 37 samples were further analysed by restriction fragment length polymorphism (RFLP) method. PCR-RFLP analysis revealed 14 strains of B. afzelii (37.8%), 15 strains of B. garinii (40.5%) and 2 strains of B. burgdorferi sensu stricto (5.4%). Four samples (10.8%) showed a mixed population of these genospecies. Two samples produced atypical RFLP pattern which were detected by sequence analysis as B. valaisiana (5.4%). Isolation attempts resulted in 21 spirochaetal strains (including two stains of B. valaisiana). The results show the diversity of B. burgdorferi s.l. in tick population and refer the risk of infection by pathogenic borreliae in Brno.
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