title: Eco-epidemiology of Borrelia miyamotoi and Lyme borreliosis spirochetes in a popular hunting and recreational forest area in Hungary.
abstract: BACKGROUND
  Borrelia miyamotoi, the newly discovered human pathogenic relapsing fever spirochete, and Borrelia burgdorferi sensu lato are maintained in natural rodent populations. The aim of this study was to investigate the natural cycle of B. miyamotoi and B. burgdorferi s.l. in a forest habitat with intensive hunting, forestry work and recreational activity in Southern Hungary.
  
  
  METHODS
  We collected rodents with modified Sherman-traps during 2010-2013 and questing ticks with flagging in 2012. Small mammals were euthanized, tissue samples were collected and all ectoparasites were removed and stored. Samples were screened for pathogens with multiplex quantitative real-time polymerase chain reaction (qPCR) targeting a part of flagellin gene, then analysed with conventional PCRs and sequencing.
  
  
  RESULTS
  177 spleen and 348 skin samples of six rodent species were individually analysed. Prevalence in rodent tissue samples was 0.2 % (skin) and 0.5 % (spleen) for B. miyamotoi and 6.6 % (skin) and 2.2 % (spleen) for B. burgdorferi s.l. Relapsing fever spirochetes were detected in Apodemus flavicollis males, B. burgdorferi s.l. in Apodemus spp. and Myodes glareolus samples. Borrelia miyamotoi was detected in one questing Ixodes ricinus nymph and B. burgdorferi s.l in nymphs and adults. In the ticks removed from rodents DNA amplification of both pathogens was successful from I. ricinus larvae (B. miyamotoi 5.6 %, B. burgdorferi s.l. 11.1 %) and one out of five nymphs while from Ixodes acuminatus larvae, and nymph only B. burgdorferi s.l. DNA was amplified. Sequencing revealed B. lusitaniae in a questing I. ricinus nymph and altogether 17 B. afzelii were identified in other samples. Two Dermacentor marginatus engorged larva pools originating from uninfected hosts were also infected with B. afzelii.
  
  
  CONCLUSIONS
  This is the first report of B. miyamotoi occurrence in a natural population of A. flavicollis as well as in Hungary. We provide new data about circulation of B. burgdorferi s.l. in rodent and tick communities including the role of I. acuminatus ticks in the endophilic pathogen cycle. Our results highlight the possible risk of infection with relapsing fever and Lyme borreliosis spirochetes in forest habitats especially in the high-risk groups of hunters, forestry workers and hikers.
authors.affiliation: Department of Parasitology and Zoology, Faculty of Veterinary Science, Szent István University, 2 István Street H-1078, Budapest, Hungary. sanyi.szekeres@gmail.com.
authors.forename: Sándor
authors.initials: S
authors.lastname: Szekeres
authors.affiliation: Laboratory for Zoonoses and Environmental Microbiology, National Institute for Public Health and Environment, 9 Antonie van Leeuwenhoeklaan, P.O. Box 1, Bilthoven, The Netherlands. claudia.coipan@rivm.nl.
authors.forename: Elena Claudia
authors.initials: EC
authors.lastname: Coipan
authors.affiliation: Department of Parasitology and Zoology, Faculty of Veterinary Science, Szent István University, 2 István Street H-1078, Budapest, Hungary. rkriszti@yahoo.com.
authors.forename: Krisztina
authors.initials: K
authors.lastname: Rigó
authors.affiliation: Department of Parasitology and Zoology, Faculty of Veterinary Science, Szent István University, 2 István Street H-1078, Budapest, Hungary. majoros.gabor@aotk.szie.hu.
authors.forename: Gábor
authors.initials: G
authors.lastname: Majoros
authors.affiliation: Laboratory for Zoonoses and Environmental Microbiology, National Institute for Public Health and Environment, 9 Antonie van Leeuwenhoeklaan, P.O. Box 1, Bilthoven, The Netherlands. seta.jahfari@rivm.nl.
authors.forename: Setareh
authors.initials: S
authors.lastname: Jahfari
authors.affiliation: Laboratory for Zoonoses and Environmental Microbiology, National Institute for Public Health and Environment, 9 Antonie van Leeuwenhoeklaan, P.O. Box 1, Bilthoven, The Netherlands. hein.sprong@rivm.nl.
authors.forename: Hein
authors.initials: H
authors.lastname: Sprong
authors.affiliation: Department of Parasitology and Zoology, Faculty of Veterinary Science, Szent István University, 2 István Street H-1078, Budapest, Hungary. FoldvariGabor@gmx.de.
authors.forename: Gábor
authors.initials: G
authors.lastname: Földvári
mesh_terms: D000818:Animals; D001898:Borrelia / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017753:Ecosystem; D065928:Forests; D006814:Hungary; D007303:Insect Vectors / Q000382:microbiology*; D060885:Multiplex Polymerase Chain Reaction; D060888:Real-Time Polymerase Chain Reaction; D012377:Rodentia / Q000469:parasitology*; D013984:Tick Infestations / Q000469:parasitology / Q000662:veterinary*; D013987:Ticks / Q000382:microbiology*