title: Isolation and biological and molecular characterization of Neospora caninum (NC-SP1) from a naturally infected adult asymptomatic cattle (Bos taurus) in the state of São Paulo, Brazil.
abstract: The biological and genetic diversity of Neospora caninum is very limited because of availability of only a few viable isolates worldwide. This study describes the isolation and biological and molecular characterization of a new viable isolate of N. caninum (NC-SP1), from a cattle in Brazil. Approximately 400 g of brain from a naturally infected adult male cattle from an abattoir was fed to a 2-month-old dog. Neospora-like oocysts were observed on day 7 post-inoculation (PI) and the duration of oocyst shedding was 14 days. The DNA obtained from oocysts was characterized molecularly and the final sequence was 99% identical to homologous sequences of N. caninum available in GenBank®. For bioassay, gerbils (Meriones unguiculatus) were orally inoculated with 10 100 and 1000 oocysts; all gerbils remained clinically normal but developed N. caninum antibodies 14 days PI. Cell culture isolation was successful using the brain homogenate from one of the gerbils and tachyzoites were observed 24 days PI. Microsatellite genotyping revealed a unique genetic profile for this new reference isolate.
authors.affiliation: Department of Preventive Veterinary Medicine and Animal Health,Faculty of Veterinary Medicine,University of São Paulo,Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo,Brazil.
authors.forename: Solange
authors.initials: S
authors.lastname: Oliveira
authors.affiliation: Department of Preventive Veterinary Medicine and Animal Health,Faculty of Veterinary Medicine,University of São Paulo,Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo,Brazil.
authors.forename: Rodrigo Martins
authors.initials: RM
authors.lastname: Soares
authors.affiliation: Department of Preventive Veterinary Medicine and Animal Health,Faculty of Veterinary Medicine,University of São Paulo,Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo,Brazil.
authors.forename: Juliana
authors.initials: J
authors.lastname: Aizawa
authors.affiliation: Department of Preventive Veterinary Medicine and Animal Health,Faculty of Veterinary Medicine,University of São Paulo,Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo,Brazil.
authors.forename: Herbert Sousa
authors.initials: HS
authors.lastname: Soares
authors.affiliation: Department of Preventive Veterinary Medicine and Animal Health,Faculty of Veterinary Medicine,University of São Paulo,Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo,Brazil.
authors.forename: Daniela Pontes
authors.initials: DP
authors.lastname: Chiebao
authors.affiliation: SALUVET, AnimalHealth Department,Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n,28040 Madrid,Spain.
authors.forename: Luis Miguel
authors.initials: LM
authors.lastname: Ortega-Mora
authors.affiliation: SALUVET, AnimalHealth Department,Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n,28040 Madrid,Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Regidor-Cerrillo
authors.affiliation: Department of Preventive Veterinary Medicine and Animal Health,Faculty of Veterinary Medicine,University of São Paulo,Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo,Brazil.
authors.forename: Natália Quadros Bressa
authors.initials: NQB
authors.lastname: Silva
authors.affiliation: Department of Preventive Veterinary Medicine and Animal Health,Faculty of Veterinary Medicine,University of São Paulo,Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo,Brazil.
authors.forename: Solange Maria
authors.initials: SM
authors.lastname: Gennari
authors.affiliation: Department of Preventive Veterinary Medicine and Animal Health,Faculty of Veterinary Medicine,University of São Paulo,Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo,Brazil.
authors.forename: Hilda Fátima Jesus
authors.initials: HFJ
authors.lastname: Pena
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D001681:Biological Assay / Q000662:veterinary; D001921:Brain / Q000469:parasitology; D001938:Brazil; D002417:Cattle; D002418:Cattle Diseases / Q000469:parasitology*; D003048:Coccidiosis / Q000469:parasitology / Q000662:veterinary*; D016054:DNA, Protozoan / Q000737:chemistry; D004285:Dogs; D005243:Feces / Q000469:parasitology; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D060005:Genotyping Techniques / Q000662:veterinary; D005849:Gerbillinae; D008297:Male; D018895:Microsatellite Repeats; D018688:Neospora / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D033661:Oocysts / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012150:Polymorphism, Restriction Fragment Length; D044967:Serum / Q000469:parasitology
chemical_list: D000913:Antibodies, Protozoan; D016054:DNA, Protozoan
keywords: Neosporosis; bioassay; cattle; gerbils; microsatellite analysis; oocyst shedding