title: Molecular detection and characterization of Anaplasma platys in dogs and ticks in Cuba.
abstract: Canine cyclic thrombocytopenia, an infectious disease caused by Anaplasma platys is a worldwide dog health problem. This study aimed to detect and characterize A. platys deoxyribonucleic acid (DNA) in dogs and ticks from Cuba using molecular methods. The study was conducted in four cities of Cuba (Habana del Este, Boyeros, Cotorro and San José de las Lajas). Blood samples were collected from 100 dogs in these cities. The animals were inspected for the detection of tick infestation and specimens were collected. Genomic DNA was extracted from dog blood and ticks using a commercial kit. Genomic DNA samples from blood and ticks were tested by a nested polymerase chain reaction (nPCR) to amplify 678 base pairs (bp) from the 16S ribosomal DNA (rDNA) of A. platys. Positive samples in nPCR were also subjected to PCR to amplify a fragment of 580bp from the citrate synthase (gltA) gene and the products were sequenced. Only Rhipicephalus sanguineus sensu lato (s.l.) was found on dogs, and 10.20% (n=5/49) of these ticks plus sixteen percent (16.0%, n=16/100) of dogs were considered positive for A. platys by nPCR targeting the 16S rDNA gene. All analyzed gltA and 16S rDNA sequences showed a 99-100% identity with sequences of A. platys reported in around the world. Phylogenetic analysis showed two defined clusters for the 16S rDNA gene and three defined clusters for the gltA gene. Based on the gltA gene, the deduced amino acid sequence showed two mutations at positions 88 and 168 compared with the sequence DQ525687 (GenBank ID from Italian sample), used as a reference in the alignment. A preliminary study on the epidemiological aspects associated with infection by A. platys showed no statistical association with the variables studied (p>0.05). This is the first evidence of the presence of A. platys in dogs and ticks in Cuba. Further studies are needed to evaluate the epidemiological aspects of A. platys infection in Cuban dogs.
authors.affiliation: Department of Animal Parasitology, Veterinary Institute, Federal Rural University of Rio de Janeiro, Br 465, km 7, Seropedica, Rio de Janeiro, 23897-000, Brazil. Electronic address: claudia_ufrrj@yahoo.com.br.
authors.forename: Claudia Bezerra da
authors.initials: CBD
authors.lastname: Silva
authors.affiliation: Department of Epidemiology and Public Health, Veterinary Institute, Federal Rural University of Rio de Janeiro, Br 465, km 7, Seropedica, Rio de Janeiro, 23897-000, Brazil.
authors.forename: Huarrisson Azevedo
authors.initials: HA
authors.lastname: Santos
authors.affiliation: Department of Animal Prevention, Veterinary Medicine College, Agrarian University of Habana, Mayabeque, Cuba.
authors.forename: Maylín González
authors.initials: MG
authors.lastname: Navarrete
authors.affiliation: Department of Animal Parasitology, Veterinary Institute, Federal Rural University of Rio de Janeiro, Br 465, km 7, Seropedica, Rio de Janeiro, 23897-000, Brazil.
authors.forename: Carla Carolina Dias Uzedo
authors.initials: CCDU
authors.lastname: Ribeiro
authors.affiliation: Department of Microbiology, National Center for Animal and Plant Health, Mayabeque, Cuba.
authors.forename: Belkis Corona
authors.initials: BC
authors.lastname: Gonzalez
authors.affiliation: Department of Microbiology, National Center for Animal and Plant Health, Mayabeque, Cuba.
authors.forename: Maykelin Fuentes
authors.initials: MF
authors.lastname: Zaldivar
authors.affiliation: Department of Animal Parasitology, Veterinary Institute, Federal Rural University of Rio de Janeiro, Br 465, km 7, Seropedica, Rio de Janeiro, 23897-000, Brazil.
authors.forename: Marcus Sandes
authors.initials: MS
authors.lastname: Pires
authors.affiliation: Department of Animal Parasitology, Veterinary Institute, Federal Rural University of Rio de Janeiro, Br 465, km 7, Seropedica, Rio de Janeiro, 23897-000, Brazil.
authors.forename: Maristela
authors.initials: M
authors.lastname: Peckle
authors.affiliation: Department of Animal Parasitology, Veterinary Institute, Federal Rural University of Rio de Janeiro, Br 465, km 7, Seropedica, Rio de Janeiro, 23897-000, Brazil.
authors.forename: Renata Lins da
authors.initials: RLD
authors.lastname: Costa
authors.affiliation: Department of Animal Parasitology, Veterinary Institute, Federal Rural University of Rio de Janeiro, Br 465, km 7, Seropedica, Rio de Janeiro, 23897-000, Brazil.
authors.forename: Gabriela Lopes Vivas
authors.initials: GLV
authors.lastname: Vitari
authors.affiliation: Department of Animal Parasitology, Veterinary Institute, Federal Rural University of Rio de Janeiro, Br 465, km 7, Seropedica, Rio de Janeiro, 23897-000, Brazil.
authors.forename: Carlos Luiz
authors.initials: CL
authors.lastname: Massard
mesh_terms: D000709:Anaplasma / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D000712:Anaplasmosis / Q000453:epidemiology* / Q000382:microbiology; D000818:Animals; D003462:Cuba / Q000453:epidemiology; D004275:DNA, Ribosomal; D004283:Dog Diseases / Q000453:epidemiology* / Q000382:microbiology; D004285:Dogs; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S; D048508:Rhipicephalus sanguineus / Q000382:microbiology*; D017422:Sequence Analysis, DNA; D011795:Surveys and Questionnaires; D013984:Tick Infestations / Q000453:epidemiology / Q000382:microbiology / Q000662:veterinary*
chemical_list: D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S
keywords: Anaplasma platys; Canine cyclic thrombocytopenia; Dogs; Polymerase chain reaction; Rhipicephalus sanguineus sensu lato