title: Characterization of two strains of Anaplasma marginale isolated from cattle in Rio de Janeiro, Brazil, after propagation in tick cell culture.
abstract: IDE8 tick cell cultures have been used for the isolation and propagation of several isolates of Anaplasma marginale. The genetic heterogeneity of A. marginale strains in cattle is diverse in endemic regions worldwide and the analyses of msp1α (major surface protein 1 alpha) gene sequences have allowed the identification of different strains. This study reports the isolation and propagation of two new isolates of A. marginale in IDE8 cells from blood of two cattle and their morphological and molecular characterization using light microscopy and the msp1α gene, respectively. Small colonies were observed in cytospin smears of each of the isolates 60 days after culture initiation. Based on msp1α sequence variation, the two isolates were found to be separate strains and were named AmRio1 and AmRio2. Analysis of msp1α microsatellite in both strains resulted in a single genotype, genotype E. The amino acid sequence of one MSP1α tandem repeat from the strain AmRio1 resulted in a new sequence (named 162) with one amino acid change. The results of these phylogenetic analyses demonstrated that A. marginale strains from Brazil and Argentina formed two large clusters of which one was less divergent that the other.
authors.affiliation: Departamento de Parasitologia Animal, Universidade Federal Rural do Rio de Janeiro, Seropédica, Rio de Janeiro, Brazil. Electronic address: babaeta@hotmail.com.
authors.forename: Bruna A
authors.initials: BA
authors.lastname: Baêta
authors.affiliation: Departamento de Parasitologia Animal, Universidade Federal Rural do Rio de Janeiro, Seropédica, Rio de Janeiro, Brazil. Electronic address: carlauzedo@yahoo.com.br.
authors.forename: Carla C D U
authors.initials: CC
authors.lastname: Ribeiro
authors.affiliation: Departamento de Parasitologia Animal, Universidade Federal Rural do Rio de Janeiro, Seropédica, Rio de Janeiro, Brazil. Electronic address: rafaella.medvet@hotmail.com.
authors.forename: Rafaella C
authors.initials: RC
authors.lastname: Teixeira
authors.affiliation: Center for Infection and Immunity of Lille (CIIL), INSERM U1019 - CNRS UMR 8204, Université Lille Nord de France, Institut Pasteur de Lille, Lille, France; SaBio, Instituto de Investigación de Recursos Cinegéticos, IREC-CSIC-UCLM-JCCM, Ciudad Real 13005, Spain. Electronic address: alejandrocabezascruz@yahoo.es.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Cabezas-Cruz
authors.affiliation: Departamento de Medicina Veterinária Preventiva, Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil. Electronic address: lygiapassos@yahoo.com.
authors.forename: Lygia M F
authors.initials: LM
authors.lastname: Passos
authors.affiliation: Institute for Comparative Tropical Medicine and Parasitology, Ludwig-Maximilians-Universität München, Munich, Germany; Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Private Bag X4, Onderstepoort 0110, South Africa.
authors.forename: Erich
authors.initials: E
authors.lastname: Zweygarth
authors.affiliation: Departamento de Parasitologia Animal, Universidade Federal Rural do Rio de Janeiro, Seropédica, Rio de Janeiro, Brazil. Electronic address: adivaldofonseca@yahoo.com.
authors.forename: Adivaldo H
authors.initials: AH
authors.lastname: Fonseca
mesh_terms: D000595:Amino Acid Sequence; D042322:Anaplasma marginale / Q000235:genetics / Q000302:isolation & purification*; D000712:Anaplasmosis / Q000382:microbiology*; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D001483:Base Sequence; D001938:Brazil; D002417:Cattle; D002418:Cattle Diseases / Q000382:microbiology*; D002460:Cell Line; D014644:Genetic Variation; D005838:Genotype; D018884:Ixodes / Q000382:microbiology*; D018895:Microsatellite Repeats / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D017422:Sequence Analysis, DNA / Q000662:veterinary; D020080:Tandem Repeat Sequences / Q000235:genetics
chemical_list: D001425:Bacterial Outer Membrane Proteins; C449089:major surface protein 1a, Anaplasma
keywords: Anaplasma marginale; Isolation; Msp1α gene; Tick cell culture