RESEARCH PAPER
Isolation, cultivation and identification of Borrelia burgdorferi genospecies from Ixodes ricinus ticks from the city of Brno, Czech Republic
 
More details
Hide details
1
Department of Anatomy, Division of Neuroanatomy, Faculty of Medicine, Masaryk University, Brno, Czech Republic
2
Department of Comparative Animal Physiology and General Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic
3
Institute of Vertebrate Biology, Academy of Sciences of the Czech Republic, Valtice, Czech Republic 4Genex CZ, Brno, Czech Republic
4
Genex CZ, Brno, Czech Republic
CORRESPONDING AUTHOR
Kateřina Pejchalová   

Department of Anatomy, Division of Neuroanatomy, Faculty of Medicine, Masaryk University, Kamenice 3, 625 00 Brno, Czech Republic.
 
Ann Agric Environ Med. 2007;14(1):75–79
KEYWORDS
ABSTRACT
A total of 305 ticks (21 larvae, 243 nymphs, 19 females and 22 males) were collectedby flagging 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-field microscopy (DFM) for the presence of borreliae. Out of 305 tick samples, 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.
 
REFERENCES (23)
1.
Anderson J: Epizootiology of Borrelia burgdorferi in Ixodes tick vector and reservoir hosts. Rev Infect Dis 1989, 11, 1451-1459.
 
2.
Bašta J, Plch J, Hulínská D, Daniel M: Incidence of Borrelia garinii and Borrelia afzelii in Ixodes ricinus ticks in an urban environment, Prague, Czech Republic, between 1995 and 1998. Eur J Clin Microbiol Infect Dis 1999, 18, 515-517.
 
3.
Burgdorfer W, Barbour AG, Hayes SF, Benach JL, Grunwaldt E, Davis JP: Lyme disease – a tick-borne spirochetosis? Science 1982, 216, 1317-1319.
 
4.
Cinco M, Padovan D, Murgia R: Detection by PCR of different species of Borrelia burgdorferi in Ixodes ricinus collected in defi ned area of northern Italy. Parassitologia 1996, 38, 378.
 
5.
Derdáková M, Beati L, Peťko B, Stanko M, Fish D: Genetic variability within Borrelia burgdorferi sensu lato genospecies established by PCR-single-strand conformation polymorfi sm analysis of the rrfA-rrlB intergenic spacer in Ixodes ricinus tick from the Czech Republic. Appl Environ Microbiol 2003, 69, 509-516.
 
6.
Gern L, Hu CM, Kociánová E, Výrosteková V, Reháček J: Genetic diversity of Borrelia burgdorferi sensu lato isolates obtained from Ixodes ricinus ticks collected in Slovakia. Eur J Epidemiol 1999, 15, 665-669.
 
7.
Hubálek Z, Korenberg EI, Juřicová Z, Kovalevskij JV, Halouzka J, Ščerbakov SV: Prevalence of Borreliae in Ixodes ricinus ticks from southern Moravia, Czechoslovakia. Folia Parasitol 1990, 37, 359-362.
 
8.
Hubálek Z, Halouzka J, Juřicová Z: A comparison of the occurrence of borreliae in nymphal and adult Ixodes ricinus ticks. Zentralbl Bakteriol 1991, 275(1), 133-137.
 
9.
Hubálek Z, Halouzka J, Juřicová Z: Prevalence of Borreliae in Ixodes ricinus ticks from urban parks. Folia Parasitol 1993, 40, 236.
 
10.
Hubálek Z, Halouzka J: Distribution of Borrelia burgdorferi sensu lato genomic groups in Europe, a review. Eur J Epidemiol 1997, 13, 951-957.
 
11.
Hubálek Z, Halouzka J: Prevalence rates of Borrelia burgdorferi sensu lato in host-seeking Ixodes ricinus ticks in Europe. Parasitol Res 1998, 84, 167-172.
 
12.
Hubálek Z, Halouzka J, Juřicová Z: Longitudinal surveillance of the tick Ixodes ricinus for Borreliae. Med Vet Entomol 2003, 17, 46-51.
 
13.
Janouškovcová E, Žákovská A. Halouzka J, Dendis M: Occurrence of Borrelia afzelii and Borrelia garinii in Ixodes ricinus ticks from southern Moravia, Czech Republic. Vector Borne Zoonotic Dis 2004, 4, 43-52.
 
14.
Kmety E, Řeháček J, Výrosteková V: Investigation of ticks for the presence of Borrelia in Czechoslovakia. Zentralbl Bakteriol 1986, 263, 468-470.
 
15.
Kurtenbach K, De Michelis S, Sewell HS, Etti S, Schafer SM, Hails R, Collares-Pereira M, Santos-Reis M, Hanincová K, Labuda M, Bormane A, Donaghy M: Distinct combinations of Borrelia burgdorferi sensu lato genospecies found in individual questing ticks from Europe. Appl Environ Microbiol 2001, 67, 4926-4929.
 
16.
O`Connell S, Granstrom M, Gray JS, Stanek G: Epidemiology of European Lyme borreliosis. Zentralbl Bakteriol 1998, 287, 229-240.
 
17.
Peťko B, Siuda K, Stanko M, Tresová G, Karbowiak G, Fričová J: Borrelia burgdorferi sensu lato in the Ixodes ricinus ticks in Southern Poland. Ann Agric Environ Med 1997, 4, 263-269.
 
18.
Schwarzová K, Čižnár I: Combined infection of Ixodes ricinus with three Borrelia burgdorferi sensu lato genotypes. Folia Microbiol 2004, 49, 297-300.
 
19.
Steere AC: Lyme disease: a growing threat to urban populations. Proc Natl Acad Sci USA, 1994, 91, 2378-2383.
 
20.
Štěpánová-Tresová G, Kopecký J, Kuthejlová M: Identifi cation of Borrelia burgdorferi sensu stricto, Borrelia garinii and Borrelia afzelii in Ixodes ricinus ticks from Southern Bohemia using monoclonal antibodies. Zentralbl Bakteriol 1999, 289, 797-806.
 
21.
Štěpánová-Tresová G, Peťko B, Štefančíková A, Nadzamová D: Occurence of Borrelia burgdorferi sensu stricto, Borrelia garinii and Borrelia afzelii in the Ixodes ricinus ticks from Eastern Slovakia. Eur J Epidemiol 2000, 16, 105-109.
 
22.
Wang G, Van Dam AP, Schwartz I, Dankert J: Molecular typing of Borrelia burgdorferi sensu lato: taxonomic, epidemiological, and clinical implications. Clin Microbiol Rev 1999, 12, 633-653.
 
23.
Žákovská A, Dendis M, Pejchalová K: Spirochaetes in Aedes species, Culex pipiens pipiens larvae and hibernating Culex pipiens molestus mosquitoes detected with dark-fi eld microscopy (DMF) and polymerase chain reaction (PCR) methods. Biológia 2000, 55, 667-670.
 
eISSN:1898-2263
ISSN:1232-1966