Molecular detection and characterization of Anaplasma platys in dogs and ticks in Cuba

被引:25
作者
da Silva, Claudia Bezerra [1 ]
Santos, Huarrisson Azevedo [2 ]
Navarrete, Maylin Gonzalez [3 ]
Dias Uzedo Ribeiro, Carla Carolina [1 ]
Gonzalez, Belkis Corona [4 ]
Zaldivar, Maykelin Fuentes [4 ]
Pires, Marcus Sandes [1 ]
Peckle, Maristela [1 ]
da Costa, Renata Lins [1 ]
Vivas Vitari, Gabriela Lopes [1 ]
Massard, Carlos Luiz [1 ]
机构
[1] Univ Fed Rural Rio de Janeiro, Dept Anim Parasitol, Vet Inst, Br 465,Km 7, BR-23897000 Rio De Janeiro, Brazil
[2] Univ Fed Rural Rio de Janeiro, Dept Epidemiol & Publ Hlth, Vet Inst, Br 465,Km 7, BR-23897000 Rio De Janeiro, Brazil
[3] Agr Univ Habana, Coll Vet Med, Dept Anim Prevent, Mayabeque, Cuba
[4] Natl Ctr Anim & Plant Hlth, Dept Microbiol, Mayabeque, Cuba
关键词
Anaplasma platys; Dogs; Canine cyclic thrombocytopenia; Polymerase chain reaction; Rhipicephalus sanguineus sensu lato; INFECTIOUS CYCLIC THROMBOCYTOPENIA; EHRLICHIA-CANIS; RHIPICEPHALUS-SANGUINEUS; DIAGNOSIS; BLOOD; PCR; IDENTIFICATION; PHAGOCYTOPHILUM; PLATELETS; SEQUENCES;
D O I
10.1016/j.ttbdis.2016.04.012
中图分类号
R51 [传染病];
学科分类号
100401 ;
摘要
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 Jose 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 580 bp 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. (C) 2016 Elsevier GmbH. All rights reserved.
引用
收藏
页码:938 / 944
页数:7
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