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Whole genome sequencing and identification of a new subtype of Borrelia lusitaniae from Ixodes ricinus tick in SloveniaKatja Strašek Smrdel,
Andraž Celar Šturm,
Alenka Babič,
Vladimir Ivović,
Eva Ružić-Sabljić, 2025, original scientific article
Abstract: Background Vector-borne diseases are becoming increasingly important in public health. The incidence of Lyme borreliosis in Slovenia continues to rise. Therefore, screening the prevalence of Borrelia spp. in vectors and their hosts is very important. Rarely, but still, it is possible to discover new species or subtypes in a new region. Methods As part of a master’s thesis, a study group collected ticks in a coastal area of Slovenia. The ticks were incubated in an MKP medium for a possible Borrelia culture. Borrelia DNA was extracted from positive cultures. We performed classical molecular genotyping methods to complement whole genome sequencing (WGS) analysis. Results Using classical genotyping, we confirmed previously undetected Borrelia species in a Slovenian tick, Borrelia lusitanie. Using large restriction fragment polymorphisms (LRFP) with the enzyme MluI, we were able to detect a new LRFP subtype of Borrelia lusitanie, namely Mll3. Using WGS, we identified a new MLST type (ST-1154) and reconstructed five linear plasmids (lp17, lp25, lp28–8, lp54 and lp56+38) and two circular plasmids (cp26 and cp32–11+1). Conclusion Regular screening of vectors and their hosts for the presence of Borrelia pathogens is important to detect the spread of different species in new areas. When reconstructing Borrelia plasmids, it is very important to apply different assembly methods and to critically analyse the results in order to select suitable contigs for further trimming and to successfully generate a complete consensus genome of high quality from hybrid read data.
Keywords: Slovenia, Borrelia lusitaniae, tick, Ixodes ricinus, WGS, plasmid analysis
Published in RUP: 05.08.2025; Views: 1219; Downloads: 14
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