title: First isolation of Neospora caninum from an aborted bovine fetus in Spain.
abstract: Neospora caninum was isolated from the brain of a 6-mo-old aborted bovine fetus from Galicia, Spain. The fetal brain homogenate was inoculated intraperitoneally into cortisonized mice. The peritoneal exudate from the infected mice, along with mouse sarcoma cells (Tg180), was inoculated into a second group of mice, and parasites were harvested from the peritoneal exudate. The parasites were adapted to in vitro growth in Vero monolayers. The tachyzoites from the peritoneal exudate reacted positively with anti-N. caninum antibodies and not with anti-Toxoplasma gondii antibodies on indirect fluorescent antibody test. The tachyzoites were lethal to interferon gamma gene knock out (KO) mice and could be identified immunohistochemically in the tissues. The identity of the parasite was also confirmed by polymerase chain reaction amplification of N. caninum-specific fragments. The sequences of the amplified gene 5 fragments (GenBank AY494944) were found to be identical to that of an Austrian isolate of N. caninum but not to that of NC-1. This is the first isolation of viable N. caninum from Spain.
authors.affiliation: CECA, Center for the Study of Animal Sciences, University of Porto, Porto, Portugal.
authors.forename: N
authors.initials: N
authors.lastname: Canada
authors.forename: C S
authors.initials: CS
authors.lastname: Meireles
authors.forename: M
authors.initials: M
authors.lastname: Mezo
authors.forename: M
authors.initials: M
authors.lastname: González-Warleta
authors.forename: J M
authors.initials: JM
authors.lastname: Correia da Costa
authors.forename: C
authors.initials: C
authors.lastname: Sreekumar
authors.forename: D E
authors.initials: DE
authors.lastname: Hill
authors.forename: K B
authors.initials: KB
authors.lastname: Miska
authors.forename: J P
authors.initials: JP
authors.lastname: Dubey
mesh_terms: D000034:Abortion, Veterinary / Q000469:parasitology*; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D001483:Base Sequence; D001921:Brain / Q000469:parasitology; D002417:Cattle; D002418:Cattle Diseases / Q000469:parasitology*; D003048:Coccidiosis / Q000469:parasitology / Q000662:veterinary*; D005260:Female; D005333:Fetus / Q000469:parasitology*; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D005849:Gerbillinae; D007371:Interferon-gamma / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D018688:Neospora / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D011247:Pregnancy; D015597:Pregnancy Complications, Parasitic / Q000276:immunology / Q000469:parasitology / Q000662:veterinary; D012689:Sequence Homology, Nucleic Acid; D013030:Spain
chemical_list: D000913:Antibodies, Protozoan; D007371:Interferon-gamma