Characterization Through Multilocus Sequence Analysis of Borrelia turdi Isolates from Portugal
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Borrelia turdi is a spirochete from the Borrelia burgdorferi complex, first reported in Japan, that has been increasingly detected in Europe. This genospecies is mostly associated with avian hosts and their ornithophilic ticks such as Ixodes frontalis. In this study, we isolated B. turdi from five I. frontalis feeding on Turdus merula, Turdus philomelos, Parus major and Troglodytes troglodytes, and one Ixodes ricinus feeding on a T. merula in Portugal. These isolates were genetically characterised according to their 5S-23S rRNA intergenic spacer, 16S rRNA and through typing of seven housekeeping genes (multilocus sequence typing). Multilocus sequence analyses revealed that the strains isolated in our study, although belonging to B. turdi genospecies, are not identical to the B. turdi reference strain Ya501. Instead, our strains are separated into a clear defined group, suggesting that the European samples diverged genetically from the strain originally detected in Japan. Population analysis of 5S-23S rRNA sequences can further resolve subpopulations within B. turdi, but more samples from a large geographical scale and host range would be needed to assess potential phylogeographical patterns within this genospecies.
KeywordsBorrelia turdi Multilocus sequence typing Isolate Portugal Ixodes frontalis Phylogenetic analysis
We would like to thank Instituto da Conservação da Natureza e Florestas, IP for logistical support, Isabel Abrantes for laboratory facilities, Pedro Lopes for help with fieldwork, Ligia Chaínho for help with laboratory work. Gabriele Margos gave valuable input regarding MLST and analysis and Dieter Heylen, Hein Sprong, Ana Palomar and José Oteo provided information on B. turdi sequences from their studies.
Ana C. Norte was financially supported by a post-doctoral fellowship from the Portuguese Foundation for Science and Technology (SFRH/BPD/62898/2009), and the study was partially funded by a centre grant (to BioISI, Centre Reference: UID/MULTI/04046/2013) from FCT/MCTES/PIDDAC, Portugal.
Compliance with Ethical Standards
The authors declare that they have no conflict of interest. All applicable international, national and institutional guidelines for the care and use of animals were followed.
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