20.500.12556/RUP-21517 Whole genome sequencing and identification of a new subtype of Borrelia lusitaniae from Ixodes ricinus tick in Slovenia 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. Slovenia Borrelia lusitaniae tick Ixodes ricinus WGS plasmid analysis Slovenija Borrelia lusitaniae klopi Ixodes ricinus WGS analiza plazmidov true false true Angleški jezik Slovenski jezik Članek v reviji 2025-08-05 09:36:44 2025-08-05 09:36:45 2025-08-06 03:04:01 0000-00-00 00:00:00 2025 0 0 str. 1-6 article 100473 Vol. 8 Sep. 2025 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 2025-07-17 579.834:595.421(497.4) 2950-1946 10.1016/j.microb.2025.100473 244826883 RAZ_Strasek_Smrdel_Katja_2025.pdf RAZ_Strasek_Smrdel_Katja_2025.pdf 1 A2CA5D3E487B6210DD3109360D62B7F2 9876735127e9bfa5cac4b739b522cffe5e92b2c01dc3f27bf17e3fef8602bc70 0e6935b8-71cf-11f0-8f0b-005056ac49c0 https://repozitorij.upr.si/Dokument.php?lang=slv&id=31288 https://www.sciencedirect.com/science/article/pii/S2950194625002419 1 f2e6be5b-71ce-11f0-8f0b-005056ac49c0 https://repozitorij.upr.si/Dokument.php?lang=slv&id=31287 Fakulteta za matematiko, naravoslovje in informacijske tehnologije 0 0 0