During the years 1996-2000, a total of 2398 Ixodes ricinus ticks were collected in three areas of southern Moravia and eastern Bohemia, Czech Republic, and examined by dark-field microscopy for the presence of spirochetes. The prevalence of B. burgdorferi sensu lato in Ixodes ricinus ticks varied and depended upon the year. In 1996, the prevalence 6.8% was observed, during 1997 and 1998, it increased to 8.4% and 12.3%, respectively. The lowest prevalence was observed in 1999 (3.6%), and in 2000 it increased to 4.0%. The mean rate of infection was 6.5%, and the proportions of infected ticks were 12.2% in 263 male ticks, 8.3% in 289 female ticks, 6.0% in 1621 nymphs, and 1.3% in 225 larvae. From the total of 156 highly infected ticks (>100 spirochetes per sample) transferred into BSK-H medium for isolation attempts, 13 isolates were obtained. PCR-RFLP and electrophoresis (SDS-PAGE) were used for the identification and characterization of Borrelia strains. Ten tick isolates were identified as Borrelia afzelii, and the other three isolates were found to be Borrelia garinii. The results indicate the epidemiological importance of B. afzelii and B. garinii in central Europe, and emphasize the role of I. ricinus in the ecology of B. burgdorferi and in the epidemiology and epizootiology of Lyme disease.

Ticks (especially those in the family Ixodidae) are the primary vectors of the infectious agent Borrelia burgdorferi sensu lato, which causes Lyme disease. To determine the potential role of mites as vectors of B. burgdorferi sensu lato, mites were collected from wild rodents in the Bazantula region of North Moravia (Czech Republic). These samples were examined for the presence of Borreliae by using DFM and PCR methods. Mites positive for the presence of DNA of B. burgdorferi sensu lato were determined as members of the families Haemogamasidae and Parasitidae. One sample from a mite of genus Haemogamasus was successfully isolated, and the specimen was confirmed as B. afzelii by using PCR-RFLP and by gradient SDS-PAGE. This suggests the possible participation of gamasid mites in borrelial circulation in nature and also points to the utility of further such studies to identify potential vectors (other than ticks) of the spirochete.

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.