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Journal of Clinical Microbiology, June 2000, p. 2128-2133, Vol. 38, No. 6
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Genetic Diversity of Borrelia burgdorferi Sensu Lato in Ticks from Mainland Portugal

Simona De Michelis,1,2 Henna-Sisko Sewell,1,2 Margarida Collares-Pereira,3 Margarida Santos-Reis,4 Leo M. Schouls,5 Vladimir Benes,6 Edward C. Holmes,1 and Klaus Kurtenbach1,2,*

The Wellcome Trust Centre for the Epidemiology of Infectious Disease, Department of Zoology, University of Oxford,1 and NERC Institute of Virology and Environmental Microbiology,2 Oxford, United Kingdom; CMDT, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa,3 and Departamento de Zoologia e Antropologia, Faculdade de Ciências, Universidade de Lisboa,4 Lisbon, Portugal; National Institute of Public Health and the Environment, Bilthoven, The Netherlands5; and European Molecular Biology Laboratory, Heidelberg, Germany6

Received 29 November 1999/Returned for modification 31 January 2000/Accepted 10 March 2000

To date Borrelia lusitaniae is the only genospecies of Borrelia burgdorferi sensu lato isolated from Ixodes ricinus ticks collected in Portugal and Tunisia. This suggests that the genospecies diversity of B. burgdorferi sensu lato decreases toward the southwestern margin of its Old World subtropical range. In order to further explore the genetic diversity of B. burgdorferi sensu lato from this region, 55 I. ricinus and 27 Hyalomma marginatum questing adults, collected during the spring of 1998 from a sylvatic habitat south of Lisbon, Portugal, were analyzed. Infection prevalences of 75% in I. ricinus ticks and 7% in H. marginatum ticks were detected by a nested PCR that targets the rrf (5S)-rrl (23S) spacer of B. burgdorferi sensu lato. Restriction fragment length polymorphism (RFLP) analysis of the I. ricinus-derived amplicons showed that the sequences in the majority of samples were similar to those of B. lusitaniae type strains (76% for strain PotiB1, 5% for strain PotiB3). Two novel RFLP patterns were obtained from 12% of the samples. The remaining 7% of samples gave mixed RFLP patterns. Phylogenetic analysis of rrf-rrl spacer sequences revealed a diverse population of B. lusitaniae in questing adult I. ricinus ticks (the sequences did not cluster with those of any other genospecies). This population consisted of 10 distinct sequence types, suggesting that multiple strains of B. lusitaniae were present in the local I. ricinus population. We hypothesize that B. lusitaniae has a narrow ecological niche that involves host species restricted to the Mediterranean Basin.

* Corresponding author. Mailing address: The Wellcome Trust Centre for the Epidemiology of Infectious Disease, Department of Zoology, University of Oxford, South Parks Rd., Oxford OX1 3PS, United Kingdom. Phone: 0044 (0)1865 281547. Fax: 0044 (0)1865 281245. E-mail: klaus.kurtenbach{at}ceid.ox.ac.uk.

Journal of Clinical Microbiology, June 2000, p. 2128-2133, Vol. 38, No. 6
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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