Isolation and molecular characterization of a new Neospora caninum isolate from cattle in Argentina.
Neospora caninum is one of the most important causes of bovine abortion, but isolation of live parasites from infected tissue is difficult. The aims of the present study were to obtain new isolates of N. caninum from congenitally infected asymptomatic newborn cattle in Argentina and to perform characterization by multilocus-microsatellite analysis. Five clinically normal born calves, with demonstrable N. caninum antibodies in precolostrum serum by indirect fluorescent antibody test, were euthanized and their brain samples were processed for histopathological, immunohistochemical, polymerase chain reaction (PCR) analysis, and for bioassay in γ-interferon knockout (GKO) mice. Although N. caninum DNA was detected in brain from all the calves by PCR, viable N. caninum was isolated in GKO mice from only one calf. Neospora caninum tachyzoites of this Argentinean isolate, designated NC-Argentina LP1, were propagated in VERO cell cultures seeded with tachyzoites from the infected GKO mice tissues. Multilocus-microsatellite typing on DNA derived from cell cultured tachyzoites revealed a unique genetic pattern, different from reported isolates. This is the first bovine isolation and genetic characterization of N. caninum in Argentina.
National Scientific and Technical Research Council (CONICET), Buenos Aires CP C1033AAJ, Argentina; Immunoparasitology Laboratory, School of Veterinary Sciences, National University of La Plata, 60 and 118, La Plata 1900, Argentina.
L M
LM
Campero
Immunoparasitology Laboratory, School of Veterinary Sciences, National University of La Plata, 60 and 118, La Plata 1900, Argentina. Electronic address: cventuri@fcv.unlp.edu.ar.
M C
MC
Venturini
National Scientific and Technical Research Council (CONICET), Buenos Aires CP C1033AAJ, Argentina.
D P
DP
Moore
National Institute of Agricultural Technology (INTA), Balcarce, 7620 Argentina.
L
L
Massola
Private Veterinary Practitioner.
H
H
Lagomarsino
National Institute of Agricultural Technology (INTA), Balcarce, 7620 Argentina.
B
B
García
Immunoparasitology Laboratory, School of Veterinary Sciences, National University of La Plata, 60 and 118, La Plata 1900, Argentina.
D
D
Bacigalupe
National Scientific and Technical Research Council (CONICET), Buenos Aires CP C1033AAJ, Argentina; Immunoparasitology Laboratory, School of Veterinary Sciences, National University of La Plata, 60 and 118, La Plata 1900, Argentina.
M
M
Rambeaud
Immunoparasitology Laboratory, School of Veterinary Sciences, National University of La Plata, 60 and 118, La Plata 1900, Argentina; Bunge and Born Foundation, Buenos Aires, Argentina.
L
L
Pardini
National Institute of Agricultural Technology (INTA), Balcarce, 7620 Argentina.
M R
MR
Leunda
Federal Research Institute for Animal Health, Institute of Epidemiology, Friedrich-Loeffler-Institute, Südufer 10, Greifswald, Insel Riems 17493, Germany.
G
G
Schares
National Institute of Agricultural Technology (INTA), Balcarce, 7620 Argentina.
C M
CM
Campero
D000818:Animals; D000831:Animals, Newborn; D000913:Antibodies, Protozoan / Q000097:blood; D001118:Argentina / Q000453:epidemiology; D001681:Biological Assay / Q000662:veterinary; D001921:Brain / Q000469:parasitology; D002417:Cattle; D002418:Cattle Diseases / Q000453:epidemiology / Q000469:parasitology*; D003048:Coccidiosis / Q000453:epidemiology / Q000469:parasitology / Q000662:veterinary*; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D007150:Immunohistochemistry / Q000662:veterinary; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D018895:Microsatellite Repeats; D008969:Molecular Sequence Data; D058885:Multilocus Sequence Typing / Q000662:veterinary; D018688:Neospora / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D016036:Seroepidemiologic Studies
D000913:Antibodies, Protozoan
Argentina; Dairy asymptomatic calf; Isolate; Multilocus-microsatellite analysis; Neospora caninum