title: Anaplasma phagocytophilum in Danish sheep: confirmation by DNA sequencing.
abstract: BACKGROUND
  The presence of Anaplasma phagocytophilum, an Ixodes ricinus transmitted bacterium, was investigated in two flocks of Danish grazing lambs. Direct PCR detection was performed on DNA extracted from blood and serum with subsequent confirmation by DNA sequencing.
  
  
  METHODS
  31 samples obtained from clinically normal lambs in 2000 from Fussingø, Jutland and 12 samples from ten lambs and two ewes from a clinical outbreak at Feddet, Zealand in 2006 were included in the study. Some of the animals from Feddet had shown clinical signs of polyarthritis and general unthriftiness prior to sampling. DNA extraction was optimized from blood and serum and detection achieved by a 16S rRNA targeted PCR with verification of the product by DNA sequencing.
  
  
  RESULTS
  Five DNA extracts were found positive by PCR, including two samples from 2000 and three from 2006. For both series of samples the product was verified as A. phagocytophilum by DNA sequencing.
  
  
  CONCLUSIONS
  A. phagocytophilum was detected by molecular methods for the first time in Danish grazing lambs during the two seasons investigated (2000 and 2006).
authors.affiliation: Department of Veterinary Disease Biology, Faculty of Life Sciences, University of Copenhagen, Dyrlaegevej 88, DK-1870 Frederiksberg C, Denmark. amkiilerich@gmail.com
authors.forename: Anne M
authors.initials: AM
authors.lastname: Kiilerich
authors.forename: Henrik
authors.initials: H
authors.lastname: Christensen
authors.forename: Stig M
authors.initials: SM
authors.lastname: Thamsborg
mesh_terms: D041081:Anaplasma phagocytophilum / Q000235:genetics*; D000818:Animals; D003718:Denmark; D016873:Ehrlichiosis / Q000175:diagnosis / Q000662:veterinary*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012756:Sheep / Q000382:microbiology; D012757:Sheep Diseases / Q000175:diagnosis* / Q000382:microbiology
chemical_list: D012336:RNA, Ribosomal, 16S