Simultaneous presence of different Borrelia burgdorferi genospecies in biological fluids of Lyme disease patients.

Oligonucleotide primers based on Borrelia burgdorferi sensu lato ospA gene sequences have been designed for use in the PCR to type all (SL primers) or each (GI to GIII primers) of the B. burgdorferi sensu lato genospecies involved in Lyme disease. These genospecies-specific primers were then used in the PCR on 24 biological fluids collected from 18 neuroborreliosis patients. Among the samples tested, 20 contained DNA from Borrelia garinii, 11 contained DNA from B. burgdorferi sensu stricto, and 10 contained DNA from Borrelia afzelii. In toto, 10 patients appeared to have been infected by a single genospecies and 8 were infected by more than one Lyme disease-associated genospecies. Serum specimens from six patients were absorbed with heterologous antigens and tested by Western blotting (immunoblotting). In four cases, residual immunodetection revealed specific epitopes of genospecies also detected by PCR; in two of them, the concordant results indicated pluri-infection of the patients. In the other two cases, Western blotting showed specific antibodies for two genospecies of Borrelia, while PCR detected DNA from only one. In summary, the data underscored the relatively high prevalence of pluri-infections in Lyme disease and confirmed the association of B. garinii with neuroborreliosis

Prevalence of <it>Borrelia burgdorferi </it>sensu lato and <it>Anaplasma phagocytophilum </it>in questing <it>Ixodes ricinus </it>ticks in relation to the density of wild cervids

<p>Abstract</p> <p>Background</p> <p><it>Borrelia burgdorferi </it>sensu lato and <it>Anaplasma phagocytophilum </it>have been considered as pathogens in animals and humans. The role of wild cervids in the epidemiology is not clear. We analyzed questing <it>Ixodes ricinus </it>ticks collected in spring for these pathogens from sites with high (Fjelløyvær and Strøm) and low density (Tjore, Hinnebu and Jomfruland) of wild cervids to study the spread of the pathogens in questing ticks.</p> <p>Methods</p> <p>For detection of <it>Anaplasma phagocytophilum </it>a 77-bp fragment in the <it>msp</it>2 gene was used. Detection of <it>Borrelia burgdorferi </it>sensu lato was performed using the FL6 and FL7 primers according to sequences of conserved regions of the <it>fla </it>gene. The <it>Osp</it>A gene located on the linear 49-kb plasmid was used as target in multiplex PCR for genotyping. Genospecies-specific primers were used in the PCR for <it>Borrelia burgdorferi </it>sensu stricto, <it>B. afzelii </it>and <it>B. garinii</it>.</p> <p>Results</p> <p>Infection rates with <it>Borrelia </it>spp. were significantly lower at Fjelløyvær and Strøm compared to Tjore and Hinnebu; Fjelløyvær vs. Tjore (χ²= 20.27, p < 0.0001); Fjelløyvær vs. Hinnebu (χ²= 24.04, p < 0.0001); Strøm vs. Tjore (χ²= 11.47, p = 0.0007) and Strøm vs. Hinnebu (χ²= 16.63, p < 0.0001). The <it>Borrelia </it>genospecies were dominated by. <it>B. afzelii </it>(82%) followed by <it>B. garinii </it>(9.7%) and <it>B. burgdorferi </it>sensu stricto (6.9%). <it>B. burgdorferi </it>s.s. was only found on the island of Jomfruland. The infection rate of <it>Anaplasma phagocytophilum </it>showed the following figures; Fjelløyvær vs Hinnebu (χ²= 16.27, p = 0.0001); Strøm vs. Tjore (χ²= 13.16, p = 0.0003); Strøm vs. Hinnebu (χ²= 34.71, p < 0.0001); Fjelløyvær vs. Tjore (χ²= 3.19, p = 0.0742) and Fjelløyvær vs. Støm (χ²= 5.06, p = 0.0245). Wild cervids may serve as a reservoir for <it>A. phagocytophilum</it>. Jomfruland, with no wild cervids but high levels of migrating birds and rodents, harboured both B. <it>burgdorferi </it>s.l. and <it>A. phagocytophilum </it>in questing <it>I. ricinus </it>ticks. Birds and rodents may play an important role in maintaining the pathogens on Jomfruland.</p> <p>Conclusion</p> <p>The high abundance of roe deer and red deer on the Norwegian islands of Fjelløyvær and Strøm may reduce the infection rate of <it>Borrelia burgdorferi </it>sensu lato in host seeking <it>Ixodes ricinus</it>, in contrast to mainland sites at Hinnebu and Tjore with moderate abundance of wild cervids. The infection rate of <it>Anaplasma phagocytophilum </it>showed the opposite result with a high prevalence in questing ticks in localities with a high density of wild cervids compared to localities with lower density.</p