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1
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Positive findings of Borrelia burgdorferi in Culex [Culex] pipiens pipiens larvae in the surrounding of Brno city determined by the PCR method

100%
Zakovska A., Nejedla P., Holikova A., Dendis M.
Annals of Agricultural and Environmental Medicine
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2002
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tom 09
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nr 2
2
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Isolation, cultivation and identification of Borrelia burgdorferi genospecies from Ixodes ricinus from the city of Brno, Czech Republic

88%
Pejchalova K., Zakovska A., Mejzlikova M., Halouzka J., Dendis M.
Annals of Agricultural and Environmental Medicine
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2007
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tom 14
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nr 1
A total of 305 ticks (21 larvae, 243 nymphs, 19 females and 22 males) were collected by fl agging of vegetation in suburban woods of Pisárky 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-fi eld 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.
3
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Isolation of Borrelia afzelii from overwintering Culex pipiens biotype Molestus mosquitoes

88%
Zakovska A., Capkova L., Sery O., Halouzka J., Dendis M.
Annals of Agricultural and Environmental Medicine
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2006
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tom 13
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nr 2
A total of 662 samples (winter period: 469; summer period: 193 specimens) of female mosquitoes of the genus Culex, Aedes and Anopheles were collected during the period March 2000-April 2001 from the locality of Vysoké Mýto (Eastern Bohemia, Czech Republic). They were examined by dark field microscopy for the presence of spirochetes. The motile spirochetes were observed in 4.2% of all species of investigated mosquitoes. One spirochetal strain out of the 8 isolation attempts (BRZ14) was obtained (cultivation rate was 12.5%) and the spirochetal strain was then successfully cultivated and identified using PCR for the presence of Borrelia burgdorferi s.l., and subsequently with the RFLP as genomospecies Borrelia afzelii. This strain was derived from overwintering Culex (Culex) pipiens biotype molestus female mosquitoe. This is apparently one of the sporadic cases of the occurrence of pathogenic borreliae in haematophagous arthropods, other than Ixodes ricinus complex ticks.
4

Identification and characterization of 31 isolates of Borrelia burgdorferi [Spirochaetales, Spirochaetaceae] obtained from various hosts and vectors using PCR-RFLP and SDS-PAGE analysis

88%
Zakovska A., Janouskovcova E., Pejchalova K., Halouzka J., Dendis M.
Acta Parasitologica
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2008
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tom 53
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nr 2
Borrelia burgdorferi sensu lato, the etiologic agent of Lyme borreliosis, circulates between ticks and vertebrate hosts. Two main genospecies typically occur in the Czech Republic Borrelia garinii and Borrelia afzelii, transmitted generally by Ixodes ricinus (L., 1758) ticks. The aim of our study was to identify spirochaete isolates focusing on Borrelia burgdorferi acquired from different sources: vectors (ticks), potential vectors (mosquitoes, small mites) and hosts (wild rodents). In the years 1996–2001 a total of 2398 ticks, 72 mites (from wild rodents), 2700 mosquito adults, 1798 mosquito larvae and organ parts (kidney and spleen) of 216 wild rodents were collected from seven localities in the Czech Republic. A total of 31 spirochaete strains were isolated: 13 strains from ticks, 1 strain from mite (Haemogamasus sp.), 15 strains from rodents, 1 strain from mosquito adults and 1 strain from mosquito larva. For the genospecies identification of these isolates PCR, PCR-RFLP was used and their characterization was also performed by SDS-PAGE. By nested PCR method all except one isolated strains were detected as Borrelia burgdorferi s.l. Following PCR-RFLP molecular analysis results, tick isolates were identified as B. garinii and B. afzelii, the strain isolated from the mite was identified as B. afzelii. This is the first isolated strain of B.b.s.l. from a different mite of infraorder Parasitiformes than tick. All of rodent isolates were identified as B. afzelii; mosquito adult isolate was identified as B. afzelii. Larval isolate from mosquito is spirochaete, but does not belong to Borrelia burgdorferi sensu lato group.
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