title: Multiple infections ofAnaplasma platysvariants in Philippine dogs.
abstract: OBJECTIVE
  Anaplasma platys, the causative agent of infectious canine cyclic thrombocytopenia, is a tick-borne pathogen that also has been implicated as potentially zoonotic. To provide molecular evidence on the multiple infections ofA. platysvariants in Philippine dogs.
  
  
  METHODS
  DNA fragments ofA. platysfrom infected dogs in the Philippines were molecularly characterized. For screening, 25 dogs suspected to have canine anaplasmosis were tested using a 16S rRNA-based nested polymerase chain reaction (PCR). Infection was confirmed by sequencing of positive amplicons. Second round PCR targeting a longer 16S rRNA fragment was subsequently performed on the first round PCR amplicons of the positive samples. Further characterization using the heat-shock operon (groEL) gene was also performed on theA. platys-positive samples.
  
  
  RESULTS
  10 16S rRNA sequences were obtained and found 99.6-100% identical to each other and 99.6-99.7% identical to the closest registeredA. platyssequences. On the other hand, 36groELclone sequences were obtained and found to be 85.1-99.8% identical with each other and 85.0-88.9% identical to the closest previously registeredA. platyssequences. Four dogs were found coinfected with 2-3groELvariant sequences. Phylogenetic analyses suggest that the detectedA. platysin the Philippines may represent unique variants.
  
  
  CONCLUSIONS
  A. platysvariants were detected in Philippine dogs. Coinfection of differentA. platysvariants in dogs was also demonstrated. The present study may indicate the potential genetic diversity ofA. platysin the country.
authors.affiliation: Department of Biology and Environmental Studies, College of Sciences, University of the Philippines Cebu, Lahug, Cebu City 6000, Philippines; Department of Research, Gullas College of Medicine, University of the Visayas, Banilad, Mandaue City 6014, Cebu, Philippines; Department of Veterinary Clinical Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Inada Cho, Hokkaido 080-8555 Japan.
authors.forename: Adrian Patalinghug
authors.initials: AP
authors.lastname: Ybañez
authors.affiliation: Department of Biology and Environmental Studies, College of Sciences, University of the Philippines Cebu, Lahug, Cebu City 6000, Philippines.
authors.forename: Rochelle Haidee Daclan
authors.initials: RH
authors.lastname: Ybañez
authors.affiliation: Research Unit for Molecular Diagnosis, National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Inada Cho, Hokkaido 080-8555, Japan.
authors.forename: Naoaki
authors.initials: N
authors.lastname: Yokoyama
authors.affiliation: Department of Veterinary Clinical Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Inada Cho, Hokkaido 080-8555 Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Inokuma
keywords: Anaplasma platys; Philippines; dogs; variants