title: Development of a pan-Babesia FRET-qPCR and a survey of livestock from five Caribbean islands.
abstract: BACKGROUND
  Babesia spp. are tick-borne protozoan hemoparasites and the second most common blood-borne parasites of mammals, in particular domestic animals. We used the Clustal Multiple Alignment program and 18S rRNA gene sequences of 22 Babesia species from GenBank to develop a PCR that could detect a wide variety of Babesia spp. in a single reaction. The pan-Babesia FRET-qPCR we developed reliably detected B. gibsoni, B. canis, B. vogeli, B. microti, B. bovis, and B. divergens under controlled conditions but did not react with closely related species, mainly Hepatozoon americanum, Theileria equi, and Toxoplasma gondii.
  
  
  RESULTS
  When we tested the pan-Babesia FRET-qPCR on DNA of whole blood from 752 cattle, sheep, goats, donkeys and horses from five Caribbean islands, we detected Babesia spp. expected to be present in the animals, mainly B. bovis and B. bigemina in cattle and B. caballi in horses and donkeys. Further, we found that animals were not uncommonly infected with species of Babesia usually associated with other hosts, mainly B. vogeli and B. gibsoni in cattle, sheep and goats, B. rossi in goats, and B. caballi in goats and sheep. Finally, the pan-Babesia FRET-qPCR enabled us to identify unknown species of Babesia in cattle, goats, sheep and donkeys.
  
  
  CONCLUSIONS
  Overall, 70 % (525/752) of the animals we tested were positive confirming earlier limited studies that infections with Babesia spp. are common in livestock in the Caribbean.
authors.affiliation: Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University College of Animal Science and Technology, Yangzhou, Jiangsu, 225009, P. R. China. lijing900401@163.com.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: Ross University School of Veterinary Medicine, Basseterre, St. Kitts and Nevis, West Indies. pkelly@rossvet.edu.kn.
authors.forename: Patrick
authors.initials: P
authors.lastname: Kelly
authors.affiliation: Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University College of Animal Science and Technology, Yangzhou, Jiangsu, 225009, P. R. China. zhangjilei0103@163.com.
authors.forename: Jilei
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University College of Animal Science and Technology, Yangzhou, Jiangsu, 225009, P. R. China. clxu@yzu.edu.cn.
authors.forename: Chuanling
authors.initials: C
authors.lastname: Xu
authors.affiliation: Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University College of Animal Science and Technology, Yangzhou, Jiangsu, 225009, P. R. China. wangcm@yzu.edu.cn.
authors.forename: Chengming
authors.initials: C
authors.lastname: Wang
mesh_terms: D000818:Animals; D001403:Babesia / Q000235:genetics / Q000302:isolation & purification*; D001404:Babesiosis / Q000175:diagnosis* / Q000453:epidemiology; D001483:Base Sequence; D016054:DNA, Protozoan / Q000235:genetics; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D058751:Livestock; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D015995:Prevalence; D016053:RNA, Protozoan; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012680:Sensitivity and Specificity; D013045:Species Specificity; D014900:West Indies / Q000453:epidemiology
chemical_list: D016054:DNA, Protozoan; D016053:RNA, Protozoan; D012337:RNA, Ribosomal, 18S