title: Detection, identification and genotyping of Borrellia spp. in rodents in Slovenia by PCR and culture.
abstract: BACKGROUND
  Borrelia burgdorferi sensu lato, the agent of Lyme borreliosis, is mainly maintained in natural foci through the transmission cycles of competent tick vectors (Ixodes sp.) and a vertebrate reservoir. Specific rodents have been identified as the principal reservoir of Borrelia burgdorferi sensu lato in Europe. Borrelia miyamotoi is the only relapsing fever spirochete transmitted by the same tick. The aim of the present study was to perform an epidemiological survey to determine the presence of B. burgdorferi sensu lato in rodents occurring in Slovenia and to explore the presence of Borrelia miyamotoi. The study was performed in two parts, retrospective and prospective; a total of 297 rodents was analyzed. Detection and identification of borrelia was performed by molecular methods and additionally in the prospective study by isolation and genotyping (MluI-LRFP and MLST).
  
  
  RESULTS
  During the prospective part of the study, borrelia was isolated from 2/46 (4.3 %) lung specimens and from 10/46 (21.7 %) heart specimens of rodents. All isolated strains were identified as B. afzelii subtype Mla1, and MLST analysis revealed 5 distinct sequence types. Borrelia DNA was successfully detected by one or other of the PCR methods in 18/46 (39.1 %) and 75/251 (29.9 %) samples in the prospective and retrospective studies, respectively. LightMix® was found to be more sensitive than the ''in-house" nested PCR (91/297 (30.6 %) vs 48/297 (16.1 %)). Borrelia miyamotoi DNA was detected in 1/251 (0.4 %) and in 1/46 (2.2 %) heart specimens, in the retrospective and prospective parts of the study, respectively.
  
  
  CONCLUSIONS
  We determined the prevalence of B. afzelii in rodents and report for the first time the presence of B. miyamotoi in Slovenia.
authors.affiliation: Faculty of Medicine, Institute of Microbiology and Immunology, University of Ljubljana, Zaloška 4, 1000, Ljubljana, Slovenia. tjasa.cerar@mf.uni-lj.si.
authors.forename: Tjaša
authors.initials: T
authors.lastname: Cerar
authors.affiliation: Faculty of Medicine, Institute of Microbiology and Immunology, University of Ljubljana, Zaloška 4, 1000, Ljubljana, Slovenia. misa.korva@mf.uni-lj.si.
authors.forename: Miša
authors.initials: M
authors.lastname: Korva
authors.affiliation: Faculty of Medicine, Institute of Microbiology and Immunology, University of Ljubljana, Zaloška 4, 1000, Ljubljana, Slovenia. tatjana.avsic@mf.uni-lj.si.
authors.forename: Tatjana
authors.initials: T
authors.lastname: Avšič-Županc
authors.affiliation: Faculty of Medicine, Institute of Microbiology and Immunology, University of Ljubljana, Zaloška 4, 1000, Ljubljana, Slovenia. eva.ruzic-sabljic@mf.uni-lj.si.
authors.forename: Eva
authors.initials: E
authors.lastname: Ružić-Sabljić
mesh_terms: D000818:Animals; D001431:Bacteriological Techniques; D001898:Borrelia / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D005838:Genotype; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D012061:Relapsing Fever / Q000453:epidemiology / Q000382:microbiology / Q000662:veterinary*; D012376:Rodent Diseases / Q000453:epidemiology / Q000382:microbiology*; D012377:Rodentia; D017524:Slovenia / Q000453:epidemiology